Stem Cell-based Embryo Models and New Perspectives for Developmental Mechanisms and Clinical Strategies

Minglei Zhi; Yixuan Yao; Chuyue Liang; Yingjie Wang; Shunxin Wang; Jeremy Kah Sheng Pang; Boon-Seng Soh; Jianyong Han

doi:10.1016/j.eng.2026.07.032

Engineering ›› :202607032 DOI: 10.1016/j.eng.2026.07.032

research-article

Stem Cell-based Embryo Models and New Perspectives for Developmental Mechanisms and Clinical Strategies

Author information +

History +

PDF (4730KB)

Abstract

Stem cell-based embryo models (SCBEMs) have emerged as important tools in developmental biology and regenerative medicine, providing novel perspectives for deciphering the mechanisms of early embryogenesis and advancing clinical applications. This review systematically summarizes recent progress in the field and proposes a three-dimensional (3D) “potency-state-lineage” classification framework that integrates the intrinsic properties of stem cells used for model construction, as well as outlining the logical framework underlying model development. This review highlights the ability of pluripotent stem cells (PSCs) to self-organize into 3D embryoids or organoids that mimic natural embryos or specific tissues. These models offer unprecedented access for studying human development while circumventing the ethical and practical constraints associated with direct research on human embryos. Moreover, these models serve as important platforms for investigating developmental disorders, environmental influences, and disease modeling. This review further explores the transformative potential of SCBEMs in personalized tissue engineering, drug screening, and livestock breeding while emphasizing the urgent need for ethical guidelines and standardization in this rapidly evolving field. SCBEM technology continues to advance and holds promise for enhancing the understanding of developmental processes and providing support for the development of therapeutic strategies.

Keywords

Stem cell-based embryo models (SCBEMs) / Pluripotent stem cells / 3D embryoid/organoid formation / Developmental biology / Regenerative medicine

Cite this article

Download citation ▾

Minglei Zhi, Yixuan Yao, Chuyue Liang, Yingjie Wang, Shunxin Wang, Jeremy Kah Sheng Pang, Boon-Seng Soh, Jianyong Han. Stem Cell-based Embryo Models and New Perspectives for Developmental Mechanisms and Clinical Strategies. Engineering 202607032 DOI:10.1016/j.eng.2026.07.032

登录浏览全文

4963

注册一个新账户忘记密码

References

Publishing order | Descend order by publishing year | Descend order by cited within

[1]	Du P, Wu J . Hallmarks of totipotent and pluripotent stem cell states. Cell Stem Cell 2024; 31(3):312-33.

[2]	Weinberger L, Ayyash M, Novershtern N, Hanna JH . Dynamic stem cell states: naive to primed pluripotency in rodents and humans. Nat Rev Mol Cell Biol 2016; 17(3):155-69.

[3]	Smith A . Formative pluripotency: the executive phase in a developmental continuum. Development 2017; 144(3):365-73.

[4]	Niakan KK, Schrode N, Cho LTY, Hadjantonakis AK . Derivation of extraembryonic endoderm stem (XEN) cells from mouse embryos and embryonic stem cells. Nat Protoc 2013; 8(6):1028-41.

[5]	Okae H, Toh H, Sato T, Hiura H, Takahashi S, Shirane K, et al. Derivation of human trophoblast stem cells. Cell Stem Cell 2018; 22(1):50-63.

[6]	Rossant J . Stem cells and early lineage development. Cell 2008; 132(4):527—31.

[7]	Macfarlan TS, Gifford WD, Driscoll S, Lettieri K, Rowe HM, Bonanomi D, et al. Embryonic stem cell potency fluctuates with endogenous retrovirus activity. Nature 2012; 487(7405):57-63.

[8]	Taubenschmid—Stowers J, Rostovskaya M, Santos F, Ljung S, Argelaguet R, Krueger F, et al. 8C—like cells capture the human zygotic genome activation program in vitro. Cell Stem Cell 2022; 29(3):449-59.

[9]	Mazid MA, Ward C, Luo Z, Liu C, Li Y, Lai Y, et al. Rolling back human pluripotent stem cells to an eight—cell embryo—like stage. Nature 2022; 605(7909):315—24.

[10]	Genet M, Torres—Padilla ME . The molecular and cellular features of 2—cell—like cells: a reference guide. Development 2020; 147(16):dev189688.

[11]	Shen H, Yang M, Li S, Zhang J, Peng B, Wang C, et al. Mouse totipotent stem cells captured and maintained through spliceosomal repression. Cell 2021; 184(11):2843-59.

[12]	Xu Y, Zhao J, Ren Y, Wang X, Lyu Y, Xie B, et al. Derivation of totipotent—like stem cells with blastocyst—like structure forming potential. Cell Res 2022; 32(6):513—29.

[13]	Rivron NC, Frias—Aldeguer J, Vrij EJ, Boisset JC, Korving J, Vivié J, et al. Blastocyst—like structures generated solely from stem cells. Nature 2018; 557(7703):106—11.

[14]	Li RH, Zhong CQ, Yu Y, Liu HS, Sakurai M, Yu LQ, et al. Generation of blastocyst—like structures from mouse embryonic and adult cell cultures. Cell 2019; 179(3):687-702.e18.

[15]	Sozen B, Cox AL, De Jonghe J, Bao M, Hollfelder F, Glover DM, et al. Self—organization of mouse stem cells into an extended potential blastoid. Dev Cell 2019; 51(6):698-712.

[16]	Yang M, Yu H, Yu X, Liang S, Hu Y, Luo Y, et al. Chemical—induced chromatin remodeling reprograms mouse ESCs to totipotent—like stem cells. Cell Stem Cell 2022; 29(3):400—18.

[17]	Hu Y, Yang Y, Tan P, Zhang Y, Han M, Yu J, et al. Induction of mouse totipotent stem cells by a defined chemical cocktail. Nature 2023; 617(7962):792—7.

[18]	Yang J, Ryan DJ, Wang W, Tsang JC, Lan G, Masaki H, et al. Establishment of mouse expanded potential stem cells. Nature 2017; 550(7676):393-7.

[19]	Yang Y, Liu B, Xu J, Wang J, Wu J, Shi C, et al. Derivation of pluripotent stem cells with in vivo embryonic and extraembryonic potency. Cell 2017; 169(2):243-57.

[20]	Posfai E, Schell JP, Janiszewski A, Rovic I, Murray A, Bradshaw B, et al. Evaluating totipotency using criteria of increasing stringency. Nat Cell Biol 2021; 23(1):49-60.

[21]	Evans MJ, Kaufman MH . Establishment in culture of pluripotential cells from mouse embryos. Nature 1981; 292(5819):154-6.

[22]	Martin GR . Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Natl Acad Sci USA 1981; 78(12):7634—8.

[23]	Boroviak T, Loos R, Bertone P, Smith A, Nichols J . The ability of inner—cell—mass cells to self—renew as embryonic stem cells is acquired following epiblast specification. Nat Cell Biol 2014; 16(6):516-28.

[24]	Boroviak T, Loos R, Lombard P, Okahara J, Behr R, Sasaki E, et al. Lineage—specific profiling delineates the emergence and progression of naive pluripotency in mammalian embryogenesis. Dev Cell 2015; 35(3):366-82.

[25]	Nichols J, Smith A . Naive and primed pluripotent states. Cell Stem Cell 2009; 4(6):487-92.

[26]	Wu J, Izpisua Belmonte JC . Dynamic pluripotent stem cell states and their applications. Cell Stem Cell 2015; 17(5):509—25.

[27]	Ying QL, Wray J, Nichols J, Batlle—Morera L, Doble B, Woodgett J, et al. The ground state of embryonic stem cell self—renewal. Nature 2008; 453(7194):519—23.

[28]	Li P, Tong C, Mehrian—Shai R, Jia L, Wu N, Yan Y, et al. Germline competent embryonic stem cells derived from rat blastocysts. Cell 2008; 135(7):1299-310.

[29]	Buehr M, Meek S, Blair K, Yang J, Ure J, Silva J, et al. Capture of authentic embryonic stem cells from rat blastocysts. Cell 2008; 135(7):1287—98.

[30]	Chen Y, Blair K, Smith A . Robust self—renewal of rat embryonic stem cells requires fine—tuning of glycogen synthase kinase—3 inhibition. Stem Cell Reports 2013; 1(3):209-17.

[31]	Chan YS, Göke J, Ng JH, Lu X, Gonzales KA, Tan CP, et al. Induction of a human pluripotent state with distinct regulatory circuitry that resembles preimplantation epiblast. Cell Stem Cell 2013; 13(6):663-75.

[32]	Gafni O, Weinberger L, Mansour AA, Manor YS, Chomsky E, Ben—Yosef D, et al. Derivation of novel human ground state naive pluripotent stem cells. Nature 2013; 504(7479):282-6.

[33]	Takashima Y, Guo G, Loos R, Nichols J, Ficz G, Krueger F, et al. Resetting transcription factor control circuitry toward ground—state pluripotency in human. Cell 2014; 158(6):1254-69.

[34]	Theunissen TW, Powell BE, Wang H, Mitalipova M, Faddah DA, Reddy J, et al. Systematic identification of culture conditions for induction and maintenance of naive human pluripotency. Cell Stem Cell 2014; 15(4):471-87.

[35]	Ware CB, Nelson AM, Mecham B, Hesson J, Zhou W, Jonlin EC, et al. Derivation of naive human embryonic stem cells. Proc Natl Acad Sci USA 2014; 111(12):4484-9.

[36]	Bayerl J, Ayyash M, Shani T, Manor YS, Gafni O, Massarwa R, et al. Principles of signaling pathway modulation for enhancing human naive pluripotency induction. Cell Stem Cell 2021; 28:1549—65.

[37]	Di Stefano B, Ueda M, Sabri S, Brumbaugh J, Huebner AJ, Sahakyan A, et al. Reduced MEK inhibition preserves genomic stability in naive human embryonic stem cells. Nat Methods 2018; 15(9):732—40.

[38]	Khan SA, Park KM, Fischer LA, Dong C, Lungjangwa T, Jimenez M, et al. Probing the signaling requirements for naive human pluripotency by high—throughput chemical screening. Cell Rep 2021; 35(11):109233.

[39]	Bredenkamp N, Yang J, Clarke J, Stirparo GG, von Meyenn F, Dietmann S, et al. Wnt inhibition facilitates RNA—mediated reprogramming of human somatic cells to naive pluripotency. Stem Cell Reports 2019; 13(6):1083-98.

[40]	Kojima Y, Kaufman—Francis K, Studdert JB, Steiner KA, Power MD, Loebel DA, et al. The transcriptional and functional properties of mouse epiblast stem cells resemble the anterior primitive streak. Cell Stem Cell 2014; 14(1):107-20.

[41]	Osorno R, Tsakiridis A, Wong F, Cambray N, Economou C, Wilkie R, et al. The developmental dismantling of pluripotency is reversed by ectopic Oct4 expression. Development 2012; 139(13):2288—98.

[42]	Tsakiridis A, Huang Y, Blin G, Skylaki S, Wymeersch F, Osorno R, et al. Distinct Wnt—driven primitive streak—like populations reflect in vivo lineage precursors. Development 2015; 142(4):809.

[43]	Thomson JA, Itskovitz—Eldor J, Shapiro SS, Waknitz MA, Swiergiel JJ, Marshall VS, et al. Embryonic stem cell lines derived from human blastocysts. Science 1998; 282(5391):1145-7.

[44]	Brons IGM, Smithers LE, Trotter MWB, Rugg—Gunn P, Sun B, Chuva de Sousa Lopes SM, et al. Derivation of pluripotent epiblast stem cells from mammalian embryos. Nature 2007; 448(7150):191—5.

[45]	Tesar PJ, Chenoweth JG, Brook FA, Davies TJ, Evans EP, Mack DL, et al. New cell lines from mouse epiblast share defining features with human embryonic stem cells. Nature 2007; 448(7150):196-9.

[46]	Xu RH, Sampsell—Barron TL, Gu F, Root S, Peck RM, Pan G, et al. NANOG is a direct target of TGFbeta/activin—mediated SMAD signaling in human ESCs. Cell Stem Cell 2008; 3(2):196-206.

[47]	Hayashi K, Ohta H, Kurimoto K, Aramaki S, Saitou M . Reconstitution of the mouse germ cell specification pathway in culture by pluripotent stem cells. Cell 2011; 146(4):519—32.

[48]	Murakami K, Günesdogan U, Zylicz JJ, Tang WWC, Sengupta R, Kobayashi T, et al. NANOG alone induces germ cells in primed epiblast in vitro by activation of enhancers. Nature 2016; 529(7586):403-7.

[49]	Ohinata Y, Ohta H, Shigeta M, Yamanaka K, Wakayama T, Saitou M . A signaling principle for the specification of the germ cell lineage in mice. Cell 2009; 137(3):571-84.

[50]	Peng B, Wang Q, Zhang F, Shen H, Du P . Mouse totipotent blastomere—like cells model embryogenesis from zygotic genome activation to post implantation. Cell Stem Cell 2025; 32(3):391-408.

[51]	Lau KYC, Rubinstein H, Gantner CW, Hadas R, Amadei G, Stelzer Y, et al. Mouse embryo model derived exclusively from embryonic stem cells undergoes neurulation and heart development. Cell Stem Cell 2022; 29(10):1445-58.e8.

[52]	Amadei G, Handford CE, Qiu C, De Jonghe J, Greenfeld H, Tran M, et al. Embryo model completes gastrulation to neurulation and organogenesis. Nature 2022; 610(7930):143-53.

[53]	Beccari L, Moris N, Girgin M, Turner DA, Baillie—Johnson P, Cossy AC, et al. Multi—axial self—organization properties of mouse embryonic stem cells into gastruloids. Nature 2018; 562(7726):272-6.

[54]	Tarazi S, Aguilera—Castrejon A, Joubran C, Ghanem N, Ashouokhi S, Roncato F, et al. Post—gastrulation synthetic embryos generated ex utero from mouse naive ESCs. Cell 2022; 185(18):3290-306.

[55]	Fan Y, Min Z, Alsolami S, Ma Z, Zhang E, Chen W, et al. Generation of human blastocyst—like structures from pluripotent stem cells. Cell Discov 2021; 7(1):81.

[56]	Sozen B, Jorgensen V, Weatherbee BAT, Chen S, Zhu M, Zernicka—Goetz M . Reconstructing aspects of human embryogenesis with pluripotent stem cells. Nat Commun 2021; 12(1):5550.

[57]	Liu LZ, Oura S, Markham Z, Hamilton JN, Skory RM, Li LJ, et al. Modeling post—implantation stages of human development into early organogenesis with stem—cell—derived peri—gastruloids. Cell 2023; 186(18):3776-92.e16.

[58]	Yanagida A, Spindlow D, Nichols J, Dattani A, Smith A, Guo G . Naive stem cell blastocyst model captures human embryo lineage segregation. Cell Stem Cell 2021; 28(6):1016-22.

[59]	Kagawa H, Javali A, Khoei HH, Sommer TM, Sestini G, Novatchkova M, et al. Human blastoids model blastocyst development and implantation. Nature 2022; 601(7894):600—5.

[60]	Yu L, Logsdon D, Pinzon—Arteaga CA, Duan J, Ezashi T, Wei Y, et al. Large—scale production of human blastoids amenable to modeling blastocyst development and maternal—fetal cross talk. Cell Stem Cell 2023; 30(9):1246-61.

[61]	Yu L, Wei Y, Duan J, Schmitz DA, Sakurai M, Wang L, et al. Blastocyst—like structures generated from human pluripotent stem cells. Nature 2021; 591(7851):620-6.

[62]	Karvas RM, Zemke JE, Ali SS, Upton E, Sane E, Fischer LA, et al. 3D—cultured blastoids model human embryogenesis from pre—implantation to early gastrulation stages. Cell Stem Cell 2023; 30(9):1148-65.

[63]	Ai Z, Niu B, Yin Y, Xiang L, Shi G, Duan K, et al. Dissecting peri—implantation development using cultured human embryos and embryo—like assembloids. Cell Res 2023; 33(9):661-78.

[64]	Oldak B, Wildschutz E, Bondarenko V, Comar MY, Zhao C, Aguilera—Castrejon A, et al. Complete human day 14 post—implantation embryo models from naive ES cells. Nature 2023; 622(7983):562-73.

[65]	Pedroza M, Gassaloglu SI, Dias N, Zhong L, Hou TJ, Kretzmer H, et al. Self—patterning of human stem cells into post—implantation lineages. Nature 2023; 622(7983):574-83.

[66]	Weatherbee BAT, Gantner CW, Iwamoto—Stohl LK, Daza RM, Hamazaki N, Shendure J, et al. Pluripotent stem cell—derived model of the post—implantation human embryo. Nature 2023; 622(7983):584-93.

[67]	Tu Z, Bi Y, Zhu X, Liu W, Hu J, Wu L, et al. Modeling human pregastrulation development by 3D culture of blastoids generated from primed—to—naïve transitioning intermediates. Protein Cell 2023; 14(5):337-49.

[68]	Shao Y, Taniguchi K, Townshend RF, Miki T, Gumucio DL, Fu J . A pluripotent stem cell—based model for post—implantation human amniotic sac development. Nat Commun 2017; 8(1):208.

[69]	Zheng Y, Xue X, Shao Y, Wang S, Esfahani SN, Li Z, et al. Controlled modelling of human epiblast and amnion development using stem cells. Nature 2019; 573(7774):421—5.

[70]	Hislop J, Song Q, Keshavarz FK, Alavi A, Schoenberger R, LeGraw R, et al. Modelling post—implantation human development to yolk sac blood emergence. Nature 2024; 626(7998):367-76.

[71]	Hamazaki N, Yang W, Kubo CA, Qiu C, Martin BK, Garge RK, et al. Retinoic acid induces human gastruloids with posterior embryo—like structures. Nat Cell Biol 2024; 26(10):1790-803.

[72]	Gharibi B, Inge OCK, Rodriguez—Hernandez I, Driscoll PC, Dubois C, Jiang M, et al. Post—gastrulation amnioids as a stem cell—derived model of human extra—embryonic development. Cell 2025; 188(14):3757-74.

[73]	Liu Y, Kim YS, Xue X, Miao Y, Kobayashi N, Sun S, et al. A human pluripotent stem cell—based somitogenesis model using microfluidics. Cell Stem Cell 2024; 31(8):1113-26.

[74]	Li J, Zhu Q, Cao J, Liu Y, Lu Y, Sun Y, et al. Cynomolgus monkey embryo model captures gastrulation and early pregnancy. Cell Stem Cell 2023; 30(4):362-77.

[75]	Pinzón—Arteaga CA, Wang Y, Wei Y, Ribeiro Orsi AE, Li L, Scatolin G, et al. Bovine blastocyst—like structures derived from stem cell cultures. Cell Stem Cell 2023; 30(5):611—16.

[76]	Xiang J, Wang H, Shi B, Li J, Liu D, Wang K, et al. Pig blastocyst—like structure models from embryonic stem cells. Cell Discov 2024; 10(1):72.

[77]	Cao J, Li J, Jin M, Shen W, Li J, Han S, et al. Sheep blastocyst—like structure models derived from stem cells. Cell Discov 2026; 12(1):20.

[78]	Kinoshita M, Barber M, Mansfield W, Cui Y, Spindlow D, Stirparo GG, et al. Capture of mouse and human stem cells with features of formative pluripotency. Cell Stem Cell 2021; 28(3):453-71.

[79]	Yu L, Wei Y, Sun HX, Mahdi AK, Pinzon Arteaga CA, Sakurai M, et al. Derivation of intermediate pluripotent stem cells amenable to primordial germ cell specification. Cell Stem Cell 2021; 28(3):550-67.

[80]	Luo Q, Pui HP, Chen J, Yu L, Jannig PR, Pei Y, et al. Epiblast—like stem cells established by Wnt/β—catenin signaling manifest distinct features of formative pluripotency and germline competence. Cell Rep 2023; 42(1):112021.

[81]	Iwatsuki K, Oikawa M, Kobayashi H, Penfold CA, Sanbo M, Yamamoto T, et al. Rat post—implantation epiblast—derived pluripotent stem cells produce functional germ cells. Cell Rep Methods 2023; 3(8):100542.

[82]	Zhi M, Zhang J, Tang Q, Yu D, Gao S, Gao D, et al. Generation and characterization of stable pig pregastrulation epiblast stem cell lines. Cell Res 2022; 32(4):383-400.

[83]	Zhi M, Gao D, Yao Y, Zhao Z, Wang Y, He P, et al. Elucidation of the pluripotent potential of bovine embryonic lineages facilitates the establishment of formative stem cell lines. Cell Mol Life Sci 2024; 81(1):427.

[84]	Zhang J, Li R, Luo R, Zhang Q, Zhao Z, Xu H, et al. Tracing and capturing the epiblast pluripotency of sheep preimplantation embryos. Adv Sci 2025; 12(36):e17764.

[85]	Zhang J, Zhi M, Gao D, Zhu Q, Gao J, Zhu G, et al. Research progress and application prospects of stable porcine pluripotent stem cells. Biol Reprod 2022; 107(1):226-36.

[86]	Zhong C, Li R, Izpisua Belmonte JC . Transcriptomic profiling fuels the derivation of stable pig epiblast stem cells. Cell Res 2022; 32(4):329—30.

[87]	Tanaka S, Kunath T, Hadjantonakis AK, Nagy A, Rossant J . Promotion of trophoblast stem cell proliferation by FGF4. Science 1998; 282(5396):2072—5.

[88]	Latos PA, Hemberger M . From the stem of the placental tree: trophoblast stem cells and their progeny. Development 2016; 143(20):3650-60.

[89]	Kunath T, Arnaud D, Uy GD, Okamoto I, Chureau C, Yamanaka Y, et al. Imprinted X—inactivation in extra—embryonic endoderm cell lines from mouse blastocysts. Development 2005; 132(7):1649-61.

[90]	Ohinata Y, Endo TA, Sugishita H, Watanabe T, Iizuka Y, Kawamoto Y, et al. Establishment of mouse stem cells that can recapitulate the developmental potential of primitive endoderm. Science 2022; 375(6580):574—8.

[91]	Dong C, Beltcheva M, Gontarz P, Zhang B, Popli P, Fischer LA, et al. Derivation of trophoblast stem cells from naïve human pluripotent stem cells. eLife 2020; 9:52504.

[92]	Io S, Kabata M, Iemura Y, Semi K, Morone N, Minagawa A, et al. Capturing human trophoblast development with naive pluripotent stem cells in vitro. Cell Stem Cell 2021; 28(6):1023—39.

[93]	Wei YX, Wang TY, Ma LS, Zhang YQ, Zhao Y, Lye K, et al. Efficient derivation of human trophoblast stem cells from primed pluripotent stem cells. Sci Adv 2021; 7(33):eabf4416.

[94]	Soncin F, Morey R, Bui T, Requena DF, Cheung VC, Kallol S, et al. Derivation of functional trophoblast stem cells from primed human pluripotent stem cells. Stem Cell Reports 2022; 17(6):1303— 17.

[95]	Xu S, Wang S, Tam TTKK, Liu P, Ruan D . Derivation of trophoblast stem cells from human expanded potential stem cells. STAR Protoc 2023; 4(2):102354.

[96]	Huang B, Peng X, Zhai X, Hu J, Chen J, Yang S, et al. Inhibition of HDAC activity directly reprograms murine embryonic stem cells to trophoblast stem cells. Dev Cell 2024; 59(16):2101—17.

[97]	Okubo T, Rivron N, Kabata M, Masaki H, Kishimoto K, Semi K, et al. Hypoblast from human pluripotent stem cells regulates epiblast development. Nature 2024; 626(7998):357—66.

[98]	Wei Y, Zhang E, Yu L, Ci B, Sakurai M, Guo L, et al. Dissecting embryonic and extraembryonic lineage crosstalk with stem cell co—culture. Cell 2023; 186(26):5859-75.

[99]	Eiraku M, Takata N, Ishibashi H, Kawada M, Sakakura E, Okuda S, et al. Self—organizing optic—cup morphogenesis in three—dimensional culture. Nature 2011; 472(7341):51-6.

[100]

Lancaster MA, Knoblich JA . Organogenesis in a dish: modeling development and disease using organoid technologies. Science 2014; 345(6194):1247125.

[101]

Sasai Y. Cytosystems dynamics in self—organization of tissue architecture. Nature 2013; 493(7432):318—26.

[102]

Clevers H . Modeling development and disease with organoids. Cell 2016; 165(7):1586-97.

[103]

Martinez Arias A, Rivron N, Moris N, Tam P, Alev C, Fu J, et al. Criteria for the standardization of stem—cell—based embryo models. Nat Cell Biol 2024; 26(10):1625-8.

[104]

Zhang HT, Hiiragi T . Symmetry breaking in the mammalian embryo. Annu Rev Cell Dev Biol 2018; 34:405-26.

[105]

Bedzhov I, Graham SJ, Leung CY, Zernicka—Goetz M . Developmental plasticity, cell fate specification and morphogenesis in the early mouse embryo. Philos Trans R Soc Lond B Biol Sci 2014; 369(1657):20130538.

[106]

Goolam M, Scialdone A, Graham SJL, Macaulay IC, Jedrusik A, Hupalowska A, et al. Heterogeneity in Oct4 and Sox2 targets biases cell fate in 4—cell mouse embryos. Cell 2016; 165(1):61-74.

[107]

Lodewijk GA, Kozuki S, Han CJ, Topacio BR, Lee S, Nixon L, et al. Self—organization of mouse embryonic stem cells into reproducible pre—gastrulation embryo models via CRISPRa programming. Cell Stem Cell 2025; 32(6):895-913.

[108]

Frum T, Murphy TM, Ralston A . HIPPO signaling resolves embryonic cell fate conflicts during establishment of pluripotency in vivo. eLife 2018; 7:e42298.

[109]

Journot RP, Huyghe M, Barthelemy A, Couto—Moreira H, Deshayes T, Harari L, et al. Conserved signals control self—organization and symmetry breaking of murine bilayered epithelia during development and regeneration. Dev Cell 2025; 60(19):2576-93.

[110]

McNamara HM, Sozen B . From genes to geometry: controlling embryo models by programming genomic activation. Cell Stem Cell 2025; 32(6):857-8.

[111]

Turing AM . The chemical basis of morphogenesis. 1953. Bull Math Biol 1990; 52(1—2):153-97.

[112]

Ferrell JE Jr. Bistability, bifurcations, and Waddington’s epigenetic landscape. Curr Biol 2012; 22(11):R458-66.

[113]

Meilhac SM, Adams RJ, Morris SA, Danckaert A, Le Garrec JF, Zernicka—Goetz M . Active cell movements coupled to positional induction are involved in lineage segregation in the mouse blastocyst. Dev Biol 2009; 331(2):210-21.

[114]

Hoffman BD, Grashoff C, Schwartz MA . Dynamic molecular processes mediate cellular mechanotransduction. Nature 2011; 475(7356):316—23.

[115]

Takeichi M . Self—organization of animal tissues: cadherin—mediated processes. Dev Cell 2011; 21(1):24-6.

[116]

Zhang S, Chen T, Chen N, Gao D, Shi B, Kong S, et al. Implantation initiation of self—assembled embryo—like structures generated using three types of mouse blastocyst—derived stem cells. Nat Commun 2019; 10(1):496.

[117]

Fu J, Warmflash A, Lutolf MP . Stem—cell—based embryo models for fundamental research and translation. Nat Mater 2021; 20(2):132-44.

[118]

Wu J, Fu J . Toward developing human organs via embryo models and chimeras. Cell 2024; 187(13):3194-219.

[119]

Grigoryan B, Paulsen SJ, Corbett DC, Sazer DW, Fortin CL, Zaita AJ, et al. Multivascular networks and functional intravascular topologies within biocompatible hydrogels. Science 2019; 364(6439):458—64.

[120]

Gómez—Álvarez M, Bueno—Fernandez C, Rodríguez—Eguren A, Francés—Herrero E, Agustina—Hernández M, Faus A, et al. Hybrid endometrial—derived hydrogels: human organoid culture models and in vivo perspectives. Adv Healthc Mater 2024; 13(11):e2303838.

[121]

Muncie JM, Ayad NME, Lakins JN, Xue X, Fu J, Weaver VM . Mechanical tension promotes formation of gastrulation—like nodes and patterns mesoderm specification in human embryonic stem cells. Dev Cell 2020; 55(6):679-94.

[122]

Tang J, Zhang P, Liu Y, Hou D, Chen Y, Cheng L, et al. Revolutionizing pressure ulcer regeneration: unleashing the potential of extracellular matrix—derived temperature—sensitive injectable antioxidant hydrogel for superior stem cell therapy. Biomaterials 2025; 314:122880.

[123]

Zhao X, Li Q, Guo Z, Li Z . Constructing a cell microenvironment with biomaterial scaffolds for stem cell therapy. Stem Cell Res Ther 2021; 12(1):583.

[124]

Virdi JK, Pethe P . Biomaterials regulate mechanosensors YAP/TAZ in stem cell growth and differentiation. Tissue Eng Regen Med 2021; 18(2):199-215.

[125]

Shahbazi MN, Pasque V . Early human development and stem cell—based human embryo models. Cell Stem Cell 2024; 31(10):1398-418.

[126]

Naahidi S, Jafari M, Logan M, Wang Y, Yuan Y, Bae H, et al. Biocompatibility of hydrogel—based scaffolds for tissue engineering applications. Biotechnol Adv 2017; 35(5):530—44.

[127]

Labouesse C, Tan BX, Agley CC, Hofer M, Winkel AK, Stirparo GG, et al. StemBond hydrogels control the mechanical microenvironment for pluripotent stem cells. Nat Commun 2021; 12(1):6132.

[128]

Gholami K, Deyhimfar R, Arefian E, Saneei Mousavi MS, Fekrirad Z, Nikoufar P, et al. Comparative characterization of hydrogels from human amniotic membrane and umbilical cord: biological and physicochemical properties. Connect Tissue Res 2025; 66(3):175-89.

[129]

Rajalekshmi R, Agrawal DK . Synergistic potential of stem cells and microfluidics in regenerative medicine. Mol Cell Biochem 2025; 480(3):1481-93.

[130]

Salazar—Ciudad I, Jernvall J, Newman SA . Mechanisms of pattern formation in development and evolution. Development 2003; 130(10):2027-37.

[131]

Schweisguth F, Corson F . Self—organization in pattern formation. Dev Cell 2019; 49(5):659-77.

[132]

Simsek MF, Özbudak EM . Patterning principles of morphogen gradients. Open Biol 2022; 12(10):220224.

[133]

Mulero—Russe A, Mora—Boza A, Marquez EN, Ziegelski M, Helmrath M, García AJ . Synthetic hydrogel substrate for human induced pluripotent stem cell definitive endoderm differentiation. Biomaterials 2025; 315:122920.

[134]

Shahbazi MN . Mechanisms of human embryo development: from cell fate to tissue shape and back. Development 2020; 147(14):dev190629.

[135]

Rossant J, Tam PPL . Early human embryonic development: blastocyst formation to gastrulation. Dev Cell 2022; 57(2):152-65.

[136]

Petropoulos S, Edsgärd D, Reinius B, Deng Q, Panula SP, Codeluppi S, et al. Single—cell RNA—seq reveals lineage and X chromosome dynamics in human preimplantation embryos. Cell 2016; 165(4):1012-26.

[137]

Brennan J, Lu CC, Norris DP, Rodriguez TA, Beddington RS, Robertson EJ . Nodal signalling in the epiblast patterns the early mouse embryo. Nature 2001; 411(6840):965-9.

[138]

Chazaud C, Yamanaka Y . Lineage specification in the mouse preimplantation embryo. Development 2016; 143(7):1063-74.

[139]

Zhai J, Xiao Z, Wang Y, Wang H . Human embryonic development: from peri—implantation to gastrulation. Trends Cell Biol 2022; 32(1):18-29.

[140]

Jia GX, Ma WJ, Wu ZB, Li S, Zhang XQ, He Z, et al. Single—cell transcriptomic characterization of sheep conceptus elongation and implantation. Cell Rep 2023; 42(8):112860.

[141]

Oestrup O, Hall V, Petkov SG, Wolf XA, Hyldig S, Hyttel P . From zygote to implantation: morphological and molecular dynamics during embryo development in the pig. Reprod Domest Anim 2009; 44(Suppl 3):39-49.

[142]

Weatherbee BAT, Weberling A, Gantner CW, Iwamoto—Stohl LK, Barnikel Z, Barrie A, et al. Distinct pathways drive anterior hypoblast specification in the implanting human embryo. Nat Cell Biol 2024; 26(3):353-65.

[143]

Aguilera—Castrejon A, Oldak B, Shani T, Ghanem N, Itzkovich C, Slomovich S, et al. Ex utero mouse embryogenesis from pre—gastrulation to late organogenesis. Nature 2021; 593(7857):119-24.

[144]

Ramos—Ibeas P, González—Brusi L, Used MT, Cocero MJ, Marigorta P, Alberio R, et al. In vitro culture of ovine embryos up to early gastrulating stages. Development 2022; 149(6):dev199743.

[145]

Bedzhov I, Leung CY, Bialecka M, Zernicka—Goetz M . In vitro culture of mouse blastocysts beyond the implantation stages. Nat Protoc 2014; 9(12):2732— 9.

[146]

Ma H, Zhai J, Wan H, Jiang X, Wang X, Wang L, et al. In vitro culture of cynomolgus monkey embryos beyond early gastrulation. Science 2019; 366(6467):eaax7890.

[147]

Niu Y, Sun N, Li C, Lei Y, Huang Z, Wu J, et al. Dissecting primate early post—implantation development using long—term in vitro embryo culture. Science 2019; 366(6467):eaaw5754.

[148]

Gantner CW, Weatherbee BAT, Wang Y, Zernicka—Goetz M . Assembly of a stem cell—derived human postimplantation embryo model. Nat Protoc 2025; 20(1):67-91.

[149]

Simunovic M, Siggia ED, Brivanlou AH . In vitro attachment and symmetry breaking of a human embryo model assembled from primed embryonic stem cells. Cell Stem Cell 2022; 29(6):962-72.

[150]

Serrano Nájera G, Weijer CJ . The evolution of gastrulation morphologies. Development 2023; 150(7):dev200885.

[151]

Sasaki K, Nakamura T, Okamoto I, Yabuta Y, Iwatani C, Tsuchiya H, et al. The germ cell fate of cynomolgus monkeys is specified in the nascent amnion. Dev Cell 2016; 39(2):169-85.

[152]

Chen D, Sun N, Hou L, Kim R, Faith J, Aslanyan M, et al. Human primordial germ cells are specified from lineage—primed progenitors. Cell Rep 2019; 29(13):4568-82.

[153]

Sheng G, Martinez Arias A, Sutherland A . The primitive streak and cellular principles of building an amniote body through gastrulation. Science 2021; 374(6572):abg1727.

[154]

Stelloo S, Alejo—Vinogradova MT, van Gelder CAGH, Zijlmans DW, van Oostrom MJ, Valverde JM, et al. Deciphering lineage specification during early embryogenesis in mouse gastruloids using multilayered proteomics. Cell Stem Cell 2024; 31(7):1072-90.

[155]

van den Brink SC, Alemany A, van Batenburg V, Moris N, Blotenburg M, Vivié J, et al. Single—cell and spatial transcriptomics reveal somitogenesis in gastruloids. Nature 2020; 582(7812):405-9.

[156]

Zhai J, Xu Y, Wan H, Yan R, Guo J, Skory R, et al. Neurulation of the cynomolgus monkey embryo achieved from 3D blastocyst culture. Cell 2023; 186:2078-91.

[157]

Yamanaka Y, Hamidi S, Yoshioka—Kobayashi K, Munira S, Sunadome K, Zhang Y, et al. Reconstituting human somitogenesis in vitro. Nature 2023; 614(7948):509—20.

[158]

Pașca SP, Arlotta P, Bateup HS, Camp JG, Cappello S, Gage FH, et al. A nomenclature consensus for nervous system organoids and assembloids. Nature 2022; 609(7929):907-10.

[159]

Xie H, An C, Bai B, Luo J, Sun N, Ci B, et al. Modeling early gastrulation in human blastoids with DNA methylation patterns of natural blastocysts. Cell Stem Cell 2025; 32(3):409-25.

[160]

Zheng Y, Yan RZ, Sun S, Kobayashi M, Xiang L, Yang R, et al. Single—cell analysis of embryoids reveals lineage diversification roadmaps of early human development. Cell Stem Cell 2022; 29(9):1402-19.

[161]

Xiang L, Yin Y, Zheng Y, Ma Y, Li Y, Zhao Z, et al. A developmental landscape of 3D—cultured human pre—gastrulation embryos. Nature 2020; 577(7791):537—42.

[162]

Shahbazi MN, Jedrusik A, Vuoristo S, Recher G, Hupalowska A, Bolton V, et al. Self—organization of the human embryo in the absence of maternal tissues. Nat Cell Biol 2016; 18(6):700-8.

[163]

Merle M, Friedman L, Chureau C, Shoushtarizadeh A, Gregor T . Precise and scalable self—organization in mammalian pseudo—embryos. Nat Struct Mol Biol 2024; 31(6):896-902.

[164]

Molè MA, Coorens THH, Shahbazi MN, Weberling A, Weatherbee BAT, Gantner CW, et al. A single cell characterisation of human embryogenesis identifies pluripotency transitions and putative anterior hypoblast centre. Nat Commun 2021; 12(1):3679.

[165]

Heidari Khoei H, Javali A, Kagawa H, Sommer TM, Sestini G, David L, et al. Generating human blastoids modeling blastocyst—stage embryos and implantation. Nat Protoc 2023; 18(5):1584-620.

[166]

Clark AT, Brivanlou A, Fu J, Kato K, Mathews D, Niakan KK, et al. Human embryo research, stem cell—derived embryo models and in vitro gametogenesis: considerations leading to the revised ISSCR guidelines. Stem Cell Reports 2021; 16(6):1416-24.

[167]

Rosa VS, Shahbazi MN . Human blastoids 3.0. Cell Stem Cell 2022; 29(1):8-10.

[168]

Shahbazi MN, Siggia ED, Zernicka—Goetz M . Self—organization of stem cells into embryos: a window on early mammalian development. Science 2019; 364(6444):948-51.

[169]

Harrison SE, Sozen B, Christodoulou N, Kyprianou C, Zernicka—Goetz M . Assembly of embryonic and extraembryonic stem cells to mimic embryogenesis in vitro. Science 2017; 356(6334):eaal1810.

[170]

Seong J, Frias—Aldeguer J, Holzmann V, Kagawa H, Sestini G, Heidari Khoei H, et al. Epiblast inducers capture mouse trophectoderm stem cells in vitro and pattern blastoids for implantation in utero. Cell Stem Cell 2022; 29(7):1102-18.

[171]

Miao Y, Djeffal Y, De Simone A, Zhu K, Lee JG, Lu Z, et al. Reconstruction and deconstruction of human somitogenesis in vitro. Nature 2023; 614(7948):500—8.

[172]

Hamazaki N, Yang W, Kubo C, Qiu C, Martin BK, Garge RK, et al. Induction and in silico staging of human gastruloids with neural tube, segmented somites & advanced cell types. 2024. bioRxiv:10.1101/2024.02.10.579769.

[173]

Veenvliet JV, Herrmann BG . Modeling mammalian trunk development in a dish. Dev Biol 2021; 474:5-15.

[174]

Wang J, Xie W, Li N, Li W, Zhang Z, Fan N, et al. Generation of a humanized mesonephros in pigs from induced pluripotent stem cells via embryo complementation. Cell Stem Cell 2023; 30(9):1235—45.

[175]

Cui J, Guo W . Establishment and long—term culture of mouse mammary stem cell organoids and breast tumor organoids. STAR Protoc 2021;2(2):100577.

[176]

Ryu S, Kim HS, Lee S, Yoon SH, Baek M, Park AY, et al. Anti—cancer drug sensitivity testing and preclinical evaluation of the anti—cancer potential of WEE1 inhibitor in triple—negative breast cancer patient—derived organoids and xenograft models. Breast Cancer Res 2025; 27(1):113.

[177]

Birtele M, Lancaster M, Quadrato G . Modelling human brain development and disease with organoids. Nat Rev Mol Cell Biol 2025; 26(5):389-412.

[178]

Yuan L, Xie S, Bai H, Liu X, Cai P, Lu J, et al. Reconstruction of dynamic mammary mini gland in vitro for normal physiology and oncogenesis. Nat Methods 2023; 20(12):2021—33.

[179]

Sahu S, Sahoo S, Sullivan T, O’Sullivan TN, Turan S, Albaugh ME, et al. Spatiotemporal modulation of growth factors directs the generation of multilineage mouse embryonic stem cell—derived mammary organoids. Dev Cell 2024; 59(2):175-86.

[180]

Xu PF, Borges RM, Fillatre J, de Oliveira—Melo M, Cheng T, Thisse B, et al. Construction of a mammalian embryo model from stem cells organized by a morphogen signalling centre. Nat Commun 2021; 12(1):3277.

[181]

Eschenhagen T, Weinberger F . Challenges and perspectives of heart repair with pluripotent stem cell—derived cardiomyocytes. Nat Cardiovasc Res 2024; 3(5):515—24.

[182]

Ryan T, Roberts JD . Stem cell models of inherited arrhythmias. Nat Cardiovasc Res 2024; 3(4):420—30.

[183]

Fiore VF, Almagro J, Fuchs E . Shaping epithelial tissues by stem cell mechanics in development and cancer. Nat Rev Mol Cell Biol 2025; 26(6):442-55.

[184]

Giulitti S, Pellegrini M, Zorzan I, Martini P, Gagliano O, Mutarelli M, et al. Direct generation of human naive induced pluripotent stem cells from somatic cells in microfluidics. Nat Cell Biol 2019; 21(2):275-86.

[185]

Kishimoto K, Iwasawa K, Sorel A, Ferran—Heredia C, Han L, Morimoto M, et al. Directed differentiation of human pluripotent stem cells into diverse organ—specific mesenchyme of the digestive and respiratory systems. Nat Protoc 2022; 17(11):2699-719.

[186]

Zheng C, Hu Y, Sakurai M, Pinzon—Arteaga CA, Li J, Wei Y, et al. Cell competition constitutes a barrier for interspecies chimerism. Nature 2021; 592(7853):272-6.

[187]

Strell P, Shetty A, Steer CJ, Low WC . Interspecies chimeric barriers for generating exogenic organs and cells for transplantation. Cell Transplant 2022; 31:9636897221110525.

[188]

Ballard E, Sakurai M, Yu L, Liu L, Oura S, Huang J, et al. Incompatibility in cell adhesion constitutes a barrier to interspecies chimerism. Cell Stem Cell 2024; 31(10):1419—26.

[189]

Huang J, He B, Wu J . Recent advances in interspecies chimeras and organogenesis. Curr Opin Genet Dev 2025; 93:102368.

[190]

Hendriks D, Pagliaro A, Andreatta F, Ma Z, van Giessen J, Massalini S, et al. Human fetal brain self—organizes into long—term expanding organoids. Cell 2024; 187(3):712-32.

[191]

Altinisik N, Rathinam D, Tran M, Gopalakrishnan J . Brain organoids restore cortical damage. Cell Stem Cell 2023; 30(3):241—2.

[192]

Fleck JS, Jansen SMJ, Wollny D, Zenk F, Seimiya M, Jain A, et al. Inferring and perturbing cell fate regulomes in human brain organoids. Nature 2023; 621(7978):365—72.

[193]

Li C, Fleck JS, Martins—Costa C, Burkard TR, Themann J, Stuempflen M, et al. Single—cell brain organoid screening identifies developmental defects in autism. Nature 2023; 621(7978):373-80.

[194]

Park DS, Kozaki T, Tiwari SK, Moreira M, Khalilnezhad A, Torta F, et al. iPS—cell—derived microglia promote brain organoid maturation via cholesterol transfer. Nature 2023; 623(7986):397-405.

[195]

Cerneckis J, Bu G, Shi Y . Pushing the boundaries of brain organoids to study Alzheimer’s disease. Trends Mol Med 2023; 29(8):659-72.

[196]

Zink A, Dai DF, Wittich A, Henke MT, Pedrotti G, Heiduschka S, et al. Pluripotent stem—cell—based screening uncovers sildenafil as a mitochondrial disease therapy. Cell 2026; 189:1656-79.

[197]

Jin J, Chen W, Li J, Yang J, Dai R, Tang J, et al. Engineered tumor microspheres via microfluidics and decellularized extracellular matrix for high—throughput organoid—based drug screening. Biofabrication 2025; 17(4):045003.

[198]

Fontanelli L, Nisini N, Pirola S, Recchia FA . Neuromuscular and cardiac organoids and assembloids: advanced platforms for drug testing. Pharmacol Ther 2025; 272:108876.

[199]

Pocock MW, Reid JD, Robinson HR, Charitakis N, Krycer JR, Foster SR, et al. Maturation of human cardiac organoids enables complex disease modeling and drug discovery. Nat Cardiovasc Res 2025; 4(7):821—40.

[200]

Xiao Y, Xu P, Bi J, Song M . Human iPSC—derived self—assembled cardiac organoids for evaluating drug developmental cardiotoxicity. Life Med 2025;4(4):lnaf018.

[201]

Li Q, Yuan Y, Zhao W, Li Y, Shi J, Xiu Y, et al. A 3D in vitro model for studying human implantation and implantation failure. Cell 2026; 189(1):70-86.

[202]

Bogliotti YS, Wu J, Vilarino M, Okamura D, Soto DA, Zhong C, et al. Efficient derivation of stable primed pluripotent embryonic stem cells from bovine blastocysts. Proc Natl Acad Sci USA 2018; 115(9):2090—5.

[203]

Zhao LX, Gao XF, Zheng YX, Wang ZX, Zhao GP, Ren J, et al. Establishment of bovine expanded potential stem cells. Proc Natl Acad Sci USA 2021; 118(15):e2018505118.

[204]

Feng R, Zhao J, Zhang Q, Zhu Z, Zhang J, Liu C, et al. Generation of anti—mastitis gene—edited dairy goats with enhancing lysozyme expression by inflammatory regulatory sequence using ISDra2—TnpB system. Adv Sci 2024; 11:e2404408.

[205]

Hou Z, An L, Han J, Yuan Y, Chen D, Tian J . Revolutionize livestock breeding in the future: an animal embryo—stem cell breeding system in a dish. J Anim Sci Biotechnol 2018; 9:90.

[206]

Mavrommatis L, Jeong HW, Kindler U, Gomez—Giro G, Kienitz MC, Stehling M, et al. Human skeletal muscle organoids model fetal myogenesis and sustain uncommitted PAX7 myogenic progenitors. eLife 2023; 12:RP87081.

[207]

Yin Y, Zhou W, Zhu J, Chen Z, Jiang L, Zhuang X, et al. Generation of self—organized neuromusculoskeletal tri—tissue organoids from human pluripotent stem cells. Cell Stem Cell 2025; 32(1):157-71.

[208]

Pereira JD, DuBreuil DM, Devlin AC, Held A, Sapir Y, Berezovski E, et al. Human sensorimotor organoids derived from healthy and amyotrophic lateral sclerosis stem cells form neuromuscular junctions. Nat Commun 2021; 12(1):4744.

[209]

Peirsman A, Nguyen HT, Van Waeyenberge M, Ceballos C, Bolivar J, Kawakita S, et al. Vascularized adipose tissue engineering: moving towards soft tissue reconstruction. Biofabrication 2023; 15(3):032003.

[210]

Strobel HA, Gerton T, Hoying JB . Vascularized adipocyte organoid model using isolated human microvessel fragments. Biofabrication 2021; 13(3):035002.

[211]

Shen JX, Couchet M, Dufau J, de Castro Barbosa T, Ulbrich MH, Helmstädter M, et al. 3D adipose tissue culture links the organotypic microenvironment to improved adipogenesis. Adv Sci 2021; 8(16):e2100106.

[212]

Faustino Martins JM, Fischer C, Urzi A, Vidal R, Kunz S, Ruffault PL, et al. Self—organizing 3D human trunk neuromuscular organoids. Cell Stem Cell 2020; 27(3):498.

[213]

Zhu G, Gao D, Li L, Yao Y, Wang Y, Zhi M, et al. Generation of three—dimensional meat—like tissue from stable pig epiblast stem cells. Nat Commun 2023; 14(1):8163.

[214]

Wu H, Wang H . Progress in stem cell—based embryo models and their applications in developmental biology and biomedicine. Nat Rev Mol Cell Biol 2026; 27(3):178-93.

[215]

Gupta A, Lutolf MP, Hughes AJ, Sonnen KF . Bioengineering in vitro models of embryonic development. Stem Cell Reports 2021; 16(5):1104— 16.

[216]

Molè MA, Elderkin S, Zorzan I, Penfold C, Horsley N, Pokhilko A, et al. Modeling human embryo implantation in vitro. Cell 2026; 189(1):87-105.

[217]

Song J, Zhao R, Zhang Y, Lu M, Liu P, Li T, et al. 3D post—implantation co—culture of human embryo and endometrium. Cell Stem Cell 2026; 33(1):58-72.

[218]

Sonnen KF . Setting the stage for embryo segmentation. Cell Stem Cell 2024; 31(8):1097—8.

[219]

Armistead FJ, Gala De Pablo J, Gadêlha H, Peyman SA, Evans SD . Cells under stress: an inertial—shear microfluidic determination of cell behavior. Biophys J 2019; 116(6):1127-35.

[220]

Xue X, Liu Y, Fu J . Bioengineering embryo models. Nat Rev Bioeng 2025; 3:11-29.

[221]

Ringel T, Frey N, Ringnalda F, Janjuha S, Cherkaoui S, Butz S, et al. Genome—scale CRISPR screening in human intestinal organoids identifies drivers of TGF—β resistance. Cell Stem Cell 2020; 26(3):431—40.

[222]

Pagliaro A, Andreatta F, Finger R, Artegiani B, Hendriks D . Generation of human fetal brain organoids and their CRISPR engineering for brain tumor modeling. Nat Protoc 2025; 20(7):1846-83.

[223]

Andreatta F, Hendriks D, Artegiani B . Human organoids as an emerging tool for genome screenings. Annu Rev Biomed Eng 2025; 27(1):157-83.

[224]

Sozen B, Amadei G, Cox A, Wang R, Na E, Czukiewska S, et al. Self—assembly of embryonic and two extra—embryonic stem cell types into gastrulating embryo—like structures. Nat Cell Biol 2018; 20(8):979-89.

[225]

Wen JH, Vincent LG, Fuhrmann A, Choi YS, Hribar KC, Taylor—Weiner H, et al. Interplay of matrix stiffness and protein tethering in stem cell differentiation. Nat Mater 2014; 13(10):979-87.

[226]

Guo J, Lyu J, Gao Z, Jia T, Wang L, Dong J, et al. 3D biomimetic niche modulates embryo development in vitro. Nat Commun 2026; 17(1):1279.

[227]

Martyn I, Kanno TY, Ruzo A, Siggia ED, Brivanlou AH . Self—organization of a human organizer by combined Wnt and Nodal signalling. Nature 2018; 558(7708):132-5.

[228]

Forristal CE, Wright KL, Hanley NA, Oreffo RO, Houghton FD . Hypoxia inducible factors regulate pluripotency and proliferation in human embryonic stem cells cultured at reduced oxygen tensions. Reproduction 2010; 139(1):85-97.

[229]

Lovell—Badge R, Anthony E, Barker RA, Bubela T, Brivanlou AH, Carpenter M, et al. ISSCR guidelines for stem cell research and clinical translation: the 2021 update. Stem Cell Reports 2021; 16(6):1398-408.

[230]

Salaris F, Rosa A . Construction of 3D in vitro models by bioprinting human pluripotent stem cells: challenges and opportunities. Brain Res 2019; 1723:146393.

[231]

Repina NA, Johnson HJ, Bao X, Zimmermann JA, Joy DA, Bi SZ, et al. Optogenetic control of Wnt signaling models cell—intrinsic embryogenic patterning using 2D human pluripotent stem cell culture. Development 2023; 150(14):dev201386.

[232]

McNamara HM, Ramm B, Toettcher JE . Synthetic developmental biology: new tools to deconstruct and rebuild developmental systems. Semin Cell Dev Biol 2023; 141:33-42.

[233]

Wang Q, Yuan F, Zuo X, Li M . Breakthroughs and challenges of organoid models for assessing cancer immunotherapy: a cutting—edge tool for advancing personalised treatments. Cell Death Discov 2025; 11(1):222.

[234]

Terhune AH, Bok J, Sun SY, Fu JP . Stem cell—based models of early mammalian development. Development 2022; 149(20):dev201015.

[235]

Hofer M, Lutolf MP . Engineering organoids. Nat Rev Mater 2021; 6(5):402-20.

[236]

Miao Y, Pek NM, Tan C, Jiang C, Yu Z, Iwasawa K, et al. Co—development of mesoderm and endoderm enables organotypic vascularization in lung and gut organoids. Cell 2025; 188(16):4295-313.

[237]

Tao B, Li X, Hao M, Tian T, Li Y, Li X, et al. Organoid—guided precision medicine: from bench to bedside. MedComm 2025; 6:e70195.

[238]

Zheng Y, Shao Y, Fu J . A microfluidics—based stem cell model of early post—implantation human development. Nat Protoc 2021; 16(1):309-26.

[239]

Jeon EY, Sorrells L, Abaci HE . Biomaterials and bioengineering to guide tissue morphogenesis in epithelial organoids. Front Bioeng Biotechnol 2022; 10:1038277.

[240]

Bai L, Wu Y, Li G, Zhang W, Zhang H, Su J . AI—enabled organoids: construction, analysis, and application. Bioact Mater 2023; 31:525—48.

[241]

Loupy A, Preka E, Chen X, Wang H, He J, Zhang K . Reshaping transplantation with AI, emerging technologies and xenotransplantation. Nat Med 2025; 31(7):2161—73.

[242]

Bao H, Zhang S, Yu Z, Xu H . Deep learning—based high—resolution time inference for deciphering dynamic gene regulation from fixed embryos. Nat Commun 2025; 16(1):6565.

[243]

Tan S, Ding Y, Wang W, Rao J, Cheng F, Zhang Q, et al. Development of an AI model for DILI—level prediction using liver organoid brightfield images. Commun Biol 2025; 8(1):886.

[244]

Iltis AS, Koster G, Reeves E, Matthews KRW . Ethical, legal, regulatory, and policy issues concerning embryoids: a systematic review of the literature. Stem Cell Res Ther 2023; 14(1):209.

[245]

Ministry of Science and Technology of the People’s Republic of China [Internet].Ethical guidelines for human organoid research. Beijing: Ministry of Science and Technology of the People’s Republic of China;2025 Apr 29 [cited 2025 Oct 10]. Available from:https://www.most.gov.cn/kjbgz/202504/P020250729547205340128.pdf.

[246]

US Department of Health and Human Services Food and Drug Administration [Internet].Guidance for Industry and Food and Drug Administration staff. Silver Spring: US Department of Health and Human Services Food and Drug Administration; 2020 July [cited 2025 Oct 10]. Available from:https://www.fda.gov/media/109176/download.

[247]

US Department of Health and Human Services Food and Drug Administration [Internet].Voluntary consensus standards recognition program for regenerative medicine therapies: guidance for industry. Silver Spring: US Department of Health and Human Services Food and Drug Administration; 2023 Oct [cited 2025 Oct 10]. Available from:https://www.fda.gov/media/159237/download.

[248]

US Department of Health and Human Services Food and Drug Administration [Internet].Recommendations for determining eligibility of donors of human cells, tissues, and cellular and tissue—based products (HCT/Ps): draft guidance for industry. Silver Spring: US Department of Health and Human Services Food and Drug Administration; 2025 Jan [cited 2025 Oct 10]. Available from:https://www.fda.gov/media/184904/download.

[249]

European Medicines Agency [Internet].Guideline on quality, non—clinical and clinical requirements for investigational advanced therapy medicinal products in clinical trials. Amsterdam: European Medicines Agency ; 2025 Apr 3 [cited 2025 Oct 10]. Available from:https://www.gmp—compliance.org/gmp—news/ema—guideline—on—quality—non—clinical—and—clinical—requirements—for—investigational—medicinal—products—for—advanced—therapies—in—clinical—trials.

[250]

European Commission [Internet].New EU rules on substances of human origin. Brussels: European Commission ; 2024 Sep 27 [cited 2025 Oct 10]. Available from:https://health.ec.europa.eu/blood—tissues—cells—and—organs/soho—regulation/new—eu—rules—substances—human—origin_en.

[251]

The European Parliament and the Council of the European Union [Internet].On advanced therapy medicinal products and amending directive 2001/83/EC and regulation (EC) No 726/2004. Brussels: the European Parliament and the Council of the European Union; 2007 Dec 10 [cited 2025 Oct 10]. Available from:https://eur—lex.europa.eu/eli/reg/2007/1394/oj/eng.