References

vavilov

Вавиловский журнал генетики и селекции

Vavilov Journal of Genetics and Breeding

2500-3259

Institute of Cytology and Genetics of Siberian Branch of the RAS

10.18699/VJ20.690

vavilov-2852

Research Article

АКТУАЛЬНЫЕ ТЕХНОЛОГИИ

MAINSTREAM TECHNOLOGIES

Использование метода бластоцистной комплементации для получения донорских органов в химерных животных

Generation of donor organs in chimeric animals via blastocyst complementation

https://orcid.org/0000-0003-3041-2789

Бабочкина

Т. И.

Babochkina

T. I.

babochkinat@yahoo.com

https://orcid.org/0000-0002-8118-1362

Герлинская

Л. А.

Gerlinskaya

L. A.

https://orcid.org/0000-0002-5388-2946

Мошкин

М. П.

Moshkin

M. P.

Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наукРоссияInstitute of Cytology and Genetics of Siberian Branch of the Russian Academy of SciencesRussian Federation

2020

31122020

248913921

2020

Бабочкина Т.И., Герлинская Л.А., Мошкин М.П.

Babochkina T.I., Gerlinskaya L.A., Moshkin M.P.

This work is licensed under a Creative Commons Attribution 4.0 License.

https://vavilov.elpub.ru/jour/article/view/2852

Сегодня актуальной проблемой в медицине является нехватка органов для трансплантаций. Одна из предполагаемых технологий получения этих органов – выращивание их из клеток человека в организме химерных животных с использованием метода межвидовой бластоцистной комплементации в комбинации с методами геномного редактирования и получения плюрипотентных стволовых клеток. Метод CRISPR/Cas9 позволяет создавать животных для бластоцистной комплементации с так называемыми свободными нишами. Совершенствование методов получения индуцированных плюрипотентных стволовых клеток дает возможность получать донорские клетки человека, способные заселять свободную нишу. Таким образом, с помощью современных технологий можно осуществить межвидовую бластоцистную комплементацию между человеком и другими животными, что в будущем позволит выращивать органы человека внутри химерных животных. Однако на практике для проведения успешной межвидовой бластоцистной комплементации необходимо решить ряд проблем: усовершенствовать методы получения «химер-компетентных клеток», преодолеть специфические межвидовые барьеры, подобрать совместимые стадии развития клеток для инъекции и соответствующего этапа развития эмбриона-реципиента, предотвратить апоптоз донорских клеток, добиться эффективной колонизации донорскими клетками человека организма животного-реципиента. Кроме того, очень важно проанализировать и законодательно урегулировать этические аспекты, возникающие при разработке технологий, связанных с получением химерных организмов с участием клеток человека. Многочисленные исследования направлены на решение этих проблем, а также на поиски новых подходов в создании межвидовых химерных организмов с целью выращивания органов человека для трансплантаций. В настоящем обзоре описаны исторические этапы развития технологии бластоцистной комплементации, детально разобраны методы, лежащие в основе ее современного варианта, и проанализированы достижения, позволяющие приблизиться к возможности выращивания органов человека в химерных животных. Рассмотрены также барьеры и проблемы, мешающие успешному применению данного подхода на практике, и дальнейшие перспективы его развития.

The lack of organs for transplantation is an important problem in medicine today. The growth of organs in chimeric animals may be the solution of this. The proposed technology is the interspecific blastocyst complementation method in combination with genomic editing for obtaining “free niches” and pluripotent stem cell production methods. The CRISPR/Cas9 method allows the so-called “free niches” to be obtained for blastocyst complementation. The technologies of producing induced pluripotent stem cells give us the opportunity to obtain human donor cells capable of populating a “free niche”. Taken together, these technologies allow interspecific blastocyst complementation between humans and other animals, which makes it possible in the future to grow human organs for transplantations inside chimeric animals. However, in practice, in order to achieve successful interspecific blastocyst complementation, it is necessary to solve a number of problems: to improve methods for producing “chimeric competent” cells, to overcome specific interspecific barriers, to select compatible cell developmental stages for injection and the corresponding developmental stage of the host embryo, to prevent apoptosis of donor cells and to achieve effective proliferation of the human donor cells in the host animal. Also, it is very important to analyze the ethical aspects related to developing technologies of chimeric organisms with the participation of human cells. Today, many researchers are trying to solve these problems and also to establish new approaches in the creation of interspecific chimeric organisms in order to grow human organs for transplantation. In the present review we described the historical stages of the development of the blastocyst complementation method, examined in detail the technologies that underlie modern blastocyst complementation, and analyzed current progress that gives us the possibility to grow human organs in chimeric animals. We also considered the barriers and issues preventing the successful implementation of interspecific blastocyst complementation in practice, and discussed the further development of this method.

химеризммежвидовой химеризмЭС клеткиИПСКCRISPR/Cas9органы для трансплантаций

chimerisminterspecies chimeraembryo SCiPSCCRISPR/Cas9organ generation

The research was supported by the Russian Science Foundation grant No. 20-14-00055, budget project No. 0324-2019-0041 and carried out using the equipment of the Center for Genetic Resources of Laboratory Animals, Federal Research Center ICG SB RAS, supported by the Ministry of Education and Science of Russia (Unique project identifier RFMEFI62119X0023)

References1

Aasen T., Raya A., Barrero M.J., Garreta E., Consiglio A., Gonzalez F., Vassena R., Bilić J., Pekarik V., Tiscornia G., Edel M., Boué S., Izpisúa Belmonte J.C. Efficient and rapid generation of induced pluripotent stem cells from human keratinocytes. Nat. Biotechnol. 2008;26(11):1276-1284. DOI 10.1038/nbt.1503.

Allegrucci C., Wu Y.Z., Thurston A., Denning C.N., Priddle H., Mummery C.L., Ward-van Oostwaard D., Andrews P.W., Stojkovic M., Smith N., Parkin T., Jones M.E., Warren G., Yu L., Brena R.M., Plass C., Young L.E. Restriction landmark genome scanning identifies culture-induced DNA methylation instability in the human embryonic stem cell epigenome. Hum. Mol. Genet. 2007;16(10):1253- 1268. DOI 10.1093/hmg/ddm074.

Barry C., Schmitz M.T., Jiang P., Schwartz M.P., Duffin B.M., Swanson S., BacherR., Bolin J.M., ElwellA.L., McIntoshB.E., StewartR., Thomson J.A. Species-specific developmental timing is maintained by pluripotent stem cells ex utero. Dev. Biol. 2017;423(2):101-110. DOI 10.1016/j.ydbio.2017.02.002.

Benchetrit H., Jaber M., Zayat V., Sebban S., Pushett A., Makedonski K., Zakheim Z., Radwan A., Maoz N., Lasry R., Renous N., Inbar M., Ram O., Kaplan T., Buganim Y. Direct induction of the three pre-implantation blastocyst cell types from fibroblasts. Cell Stem Cell. 2019;24(6):983-994.e7. DOI 10.1016/j.stem.2019.03.018.

Betschinger J., Nichols J., Dietmann S., Corrin P.D., Paddison P.J., Smith A. Exit from pluripotency is gated by intracellular redistribution of the bHLH transcription factor Tfe3. Cell. 2013;153(2):335- 347. DOI 10.1016/j.cell.2013.03.012.

Boroviak K., Doe B., Banerjee R., Yang F., Bradley A. Chromosome engineering in zygotes with CRISPR/Cas9. Genesis. 2016;54(2): 78-85. DOI 10.1002/dvg.22915.

Bourret R., Martinez E., Vialla F., Giquel C., Thonnat-Marin A., De Vos J. Human-animal chimeras: ethical issues about farming chimeric animals bearing human organs. Stem Cell Res. Ther. 2016;7(1): 87. DOI 10.1186/s13287-016-0345-9.

Bradley A., Evans M., Kaufman M.H., Robertson E. Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines. Nature. 1984;309(5965):255-256. DOI 10.1038/309255a0.

Carbery I.D., Ji D., Harrington A., Brown V., Weinstein E.J., Liaw L., Cui X. Targeted genome modification in mice using zinc-finger nucleases. Genetics. 2010;186(2):451-459. DOI 10.1534/genetics.110.117002.

Chang A.N., Liang Z., Dai H.Q., Chapdelaine-Williams A.M., Andrews N., Bronson R.T., Schwer B., Alt F.W. Neural blastocyst complementation enables mouse forebrain organogenesis. Nature. 2018; 563(7729):126-130. DOI 10.1038/s41586-018-0586-0.

Chen J., Lansford R., Stewart V., Young F., Alt F.W. RAG-2-deficient blastocyst complementation: an assay of gene function in lymphocyte development. Proc. Natl. Acad. Sci. USA. 1993;90(10):4528- 4532. DOI 10.1073/pnas.90.10.4528.

Chen K., Chmait R.H., Vanderbilt D., Wu S., Randolph L. Chimerism in monochorionic dizygotic twins: case study and review. Am. J. Med. Genet. A. 2013;161A(7):1817-1824. DOI 10.1002/ajmg.a.35957.

Chen Y., Niu Y., Li Y., Ai Z., Kang Y., Shi H., Xiang Z., Yang Z., Tan T., Si W., Li W., Xia X., Zhou Q., Ji W., Li T. Generation of cynomolgus monkey chimeric fetuses using embryonic stem cells. Cell Stem Cell. 2015;17(1):116-124. DOI 10.1016/j.stem.2015.06.004.

Cong L., Ran F.A., Cox D., Lin S., Barretto R., Habib N., Hsu P.D., Wu X., Jiang W., Marraffini L.A., Zhang F. Multiplex genome engineering using CRISPR/Cas systems. Science. 2013;339(6121):819- 823. DOI 10.1126/science.1231143.

De Los Angeles A., Elsworth J.D., Redmond D.E. ERK-independent African Green monkey pluripotent stem cells in a putative chimeracompetent state. Biochem. Biophys. Res. Commun. 2019;510(1):78- 84. DOI 10.1016/j.bbrc.2019.01.037.

Drexler C., Glock B., Vadon M., Staudacher E., Dauber E.M., Ulrich S., Reisacher B.K., Mayr W.R., Lanzer G., Wagner T. Tetragametic chimerism detected in a healthy woman with mixed-field agglutination reactions in ABO blood grouping. Transfusion. 2005;45(5):698-703. DOI 10.1111/j.1537-2995.2005.04304.x.

Dunn S.J., Martello G., Yordanov B., Emmott S., Smith A.G. Defining an essential transcription factor program for naïve pluripotency. Science. 2014;344(6188):1156-1160. DOI 10.1126/science.1248882.

Evans M.J., Kaufman M.H. Establishment in culture of pluripotential cells from mouse embryos. Nature. 1981;292(5819):154-156. DOI 10.1038/292154a.

Fang R., Liu K., Zhao Y., Li H., Zhu D., Du Y., Xiang C., Li X., Liu H., Miao Z., Zhang X., Shi Y., Yang W., Xu J., Deng H. Generation of naive induced pluripotent stem cells from rhesus monkey fibroblasts. Cell Stem Cell. 2014;15(4):488-497. DOI 10.1016/j.stem.2014.09.004.

Farahany N.A., Greely H.T., Hyman S., Koch C., Grady C., Pașca S.P., Sestan N., Arlotta P., Bernat J.L., Ting J., Lunshof J.E., Iyer E., Hyun I., Capestany B.H., Church G.M., Huang H., Song H. The ethics of experimenting with human brain tissue. Nature. 2018; 556(7702):429-432. DOI 10.1038/d41586-018-04813-x.

Friel R., van der Sar S., Mee P.J. Embryonic stem cells: understanding their history, cell biology and signalling. Adv. Drug. Deliv. Rev. 2005;57(13):1894-1903. DOI 10.1016/j.addr.2005.08.002.

Fu R., Yu D., Ren J., Li C., Wang J., Feng G., Wang X., Wan H., Li T., Wang L., Zhang Y., Hai T., Li W., Zhou Q. Domesticated cynomolgus monkey embryonic stem cells allow the generation of neonatal interspecies chimeric pigs. Protein Cell. 2020;11(2):97-107. DOI 10.1007/s13238-019-00676-8.

Gafni O., Weinberger L., Mansour A.A., Manor Y.S., Chomsky E., BenYosef D., Kalma Y., Viukov S., Maza I., Zviran A., Rais Y., Shipony Z., Mukamel Z., Krupalnik V., Zerbib M., Geula S., Caspi I., Schneir D., Shwartz T., Gilad S., Amann-Zalcenstein D., Benjamin S., Amit I., Tanay A., Massarwa R., Novershtern N., Hanna J.H. Derivation of novel human ground state naive pluripotent stem cells. Nature. 2013;504(7479):282-286. DOI 10.1038/nature12745.

Gardner R.L. Mouse chimeras obtained by the injection of cells into the blastocyst. Nature. 1968;220(5167):596-597. DOI 10.1038/220596a0.

Goto T., Hara H., Sanbo M., Masaki H., Sato H., Yamaguchi T., Hochi S., Kobayashi T., Nakauchi H., Hirabayashi M. Generation of pluripotent stem cell-derived mouse kidneys in Sall1-targeted anephric rats. Nat. Commun. 2019;10(1):451. DOI 10.1038/s41467-019-08394-9.

Guo G., Yang J., Nichols J., Hall J.S., Eyres I., Mansfield W., Smith A. Klf4 reverts developmentally programmed restriction of ground state pluripotency. Development. 2009;136(7):1063-1069. DOI 10.1242/dev.030957.

Hamanaka S., Umino A., Sato H., Hayama T., Yanagida A., Mizuno N., Kobayashi T., Kasai M., Suchy F.P., Yamazaki S., Masaki H., Yamaguchi T., Nakauchi H. Generation of vascular endothelial cells and hematopoietic cells by blastocyst complementation. Stem Cell Reports. 2018;11(4):988-997. DOI 10.1016/j.stemcr.2018.08.015.

Hanna J., Cheng A.W., Saha K., Kim J., Lengner C.J., Soldner F., Cassady J.P., Muffat J., Carey B.W., Jaenisch R. Human embryonic stem cells with biological and epigenetic characteristics similar to those of mouse ESCs. Proc. Natl. Acad. Sci. USA. 2010;107(20):9222- 9227. DOI 10.1073/pnas.1004584107.

Harrison S.E., Sozen B., Christodoulou N., Kyprianou C., ZernickaGoetz M. Assembly of embryonic and extraembryonic stem cells to mimic embryogenesis in vitro. Science. 2017;356(6334). DOI 10.1126/science.aal1810.

Hashimoto H., Eto T., Yamamoto M., Yagoto M., Goto M., Kagawa T., Kojima K., Kawai K., Akimoto T., Takahashi R.I. Development of blastocyst complementation technology without contributions to gametes and the brain. Exp. Anim. 2019;68(3):361-370. DOI 10.1538/expanim.18-0173.

Hayashi K., Hikabe O., Obata Y., Hirao Y. Reconstitution of mouse oogenesis in a dish from pluripotent stem cells. Nat. Protoc. 2017; 12(9):1733-1744. DOI 10.1038/nprot.2017.070.

Hu Z., Li H., Jiang H., Ren Y., Yu X., Qiu J., Stablewski A.B., Zhang B., Buck M.J., Feng J. Transient inhibition of mTOR in human pluripotent stem cells enables robust formation of mouse-human chimeric embryos. Sci. Adv. 2020;6(20):eaaz0298. DOI 10.1126/sciadv.aaz0298.

Huang Y., Liang P., Liu D., Huang J., Songyang Z. Telomere regulation in pluripotent stem cells. Protein Cell. 2014;5(3):194-202. DOI 10.1007/s13238-014-0028-1.

Huang Y., Osorno R., Tsakiridis A., Wilson V. In vivo differentiation potential of epiblast stem cells revealed by chimeric embryo formation. Cell Rep. 2012;2(6):1571-1578. DOI 10.1016/j.celrep.2012.10.022.

Isotani A., Hatayama H., Kaseda K., Ikawa M., Okabe M. Formation of a thymus from rat ES cells in xenogeneic nude mouse↔rat ES chimeras. Genes Cells. 2011;16(4):397-405. DOI 10.1111/j.1365-2443.2011.01495.x.

James D., Noggle S.A., Swigut T., Brivanlou A.H. Contribution of human embryonic stem cells to mouse blastocysts. Dev. Biol. 2006; 295(1):90-102. DOI 10.1016/j.ydbio.2006.03.026.

Kilens S., Meistermann D., Moreno D., Chariau C., Gaignerie A., Reignier A., Lelièvre Y., Casanova M., Vallot C., Nedellec S., Flippe L., Firmin J., Song J., Charpentier E., Lammers J., Donnart A., Marec N., Deb W., Bihouée A., Le Caignec C., Pecqueur C., Redon R., Barrière P., Bourdon J., Pasque V., Soumillon M., Mikkelsen T.S., Rougeulle C., Fréour T., David L., Milieu Intérieur. Consortium. Parallel derivation of isogenic human primed and naive induced pluripotent stem cells. Nat. Commun. 2018;9(1):360. DOI 10.1038/s41467-017-02107-w.

Kobayashi T., Yamaguchi T., Hamanaka S., Kato-Itoh M., Yamazaki Y., Ibata M., Sato H., Lee Y.S., Usui J., Knisely A.S., Hirabayashi M., Nakauchi H. Generation of rat pancreas in mouse by interspecific blastocyst injection of pluripotent stem cells. Cell. 2010;142(5):787- 799. DOI 10.1016/j.cell.2010.07.039.

Liu X., Nefzger C.M., Rossello F.J., Chen J., Knaupp A.S., Firas J., Ford E., Pflueger J., Paynter J.M., Chy H.S., O’Brien C.M., Huang C., Mishra K., Hodgson-Garms M., Jansz N., Williams S.M., Blewitt M.E., Nilsson S.K., Schittenhelm R.B., Laslett A.L., Lister R., Polo J.M. Comprehensive characterization of distinct states of human naive pluripotency generated by reprogramming. Nat. Methods. 2017;14(11):1055-1062. DOI 10.1038/nmeth.4436.

MacLaren L.A., Anderson G.B., BonDurant R.H., Edmondson A.J. Inter- and intraspecific placentae in sheep, goats and sheep-goat chimaeras. J. Comp. Pathol. 1992;106(3):279-297. DOI 10.1016/ 0021-9975(92)90056-z.

Mali P., Yang L., Esvelt K.M., Aach J., Guell M., DiCarlo J.E., Norville J.E., Church G.M. RNA-guided human genome engineering via Cas9. Science. 2013;339(6121):823-826. DOI 10.1126/science.1232033.

Martin G.R. 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):7634-7638. DOI 10.1073/pnas.78.12.7634.

Masaki H., Kato-Itoh M., Takahashi Y., Umino A., Sato H., Ito K., Yanagida A., Nishimura T., Yamaguchi T., Hirabayashi M., Era T., Loh K.M., Wu S.M., Weissman I.L., Nakauchi H. Inhibition of apoptosis overcomes stage-related compatibility barriers to chimera formation in mouse embryos. Cell Stem Cell. 2016;19(5):587-592. DOI 10.1016/j.stem.2016.10.013.

Mascetti V.L., Pedersen R.A. Contributions of mammalian chimeras to pluripotent stem cell research. Cell Stem Cell. 2016a;19(2):163-175. DOI 10.1016/j.stem.2016.07.018.

Mascetti V.L., Pedersen R.A. Human-mouse chimerism validates human stem cell pluripotency. Cell Stem Cell. 2016b;18(1):67-72. DOI 10.1016/j.stem.2015.11.017.

Matsunari H., Watanabe M., Hasegawa K., Uchikura A., Nakano K., Umeyama K., Masaki H., Hamanaka S., Yamaguchi T., Nagaya M., Nishinakamura R., Nakauchi H., Nagashima H. Compensation of disabled organogeneses in genetically modified pig fetuses by blastocyst complementation. Stem Cell Reports. 2020;14(1):21-33. DOI 10.1016/j.stemcr.2019.11.008.

McLaren A., Bowman P. Mouse chimaeras derived from fusion of embryos differing by nine genetic factors. Nature. 1969;224(5216): 238-240. DOI 10.1038/224238a0.

Mintz B. Genetic mosaicism in adult mice of quadriparental lineage. Science. 1965;148(3674):1232-1233. DOI 10.1126/science.148.3674.1232.

Nelson J.L., Furst D.E., Maloney S., Gooley T., Evans P.C., Smith A., Bean M.A., Ober C., Bianchi D.W. Microchimerism and HLA-compatible relationships of pregnancy in scleroderma. Lancet. 1998; 351(9102):559-562. DOI 10.1016/S0140-6736(97)08357-8.

Nichols J., Smith A. Naive and primed pluripotent states. Cell Stem Cell. 2009;4(6):487-492. DOI 10.1016/j.stem.2009.05.015.

Offield M.F., Jetton T.L., Labosky P.A., Ray M., Stein R.W., Magnuson M.A., Hogan B.L., Wright C.V. PDX-1 is required for pancreatic outgrowth and differentiation of the rostral duodenum. Development. 1996;122(3):983-995.

Ohinata Y., Payer B., O’Carroll D., Ancelin K., Ono Y., Sano M., Barton S.C., Obukhanych T., Nussenzweig M., Tarakhovsky A., Saitou M., Surani M.A. Blimp1 is a critical determinant of the germ cell lineage in mice. Nature. 2005;436(7048):207-213. DOI 10.1038/nature03813.

Okita K., Ichisaka T., Yamanaka S. Generation of germline-competent induced pluripotent stem cells. Nature. 2007;448(7151):313-317. DOI 10.1038/nature05934.

Okumura H., Nakanishi A., Toyama S., Yamanoue M., Yamada K., Ukai A., Hashita T., Iwao T., Miyamoto T., Tagawa Y.I., Hirabayashi M., Miyoshi I., Matsunaga T. Contribution of rat embryonic stem cells to xenogeneic chimeras in blastocyst or 8-cell embryo injection and aggregation. Xenotransplantation. 2019;26(1):e12468. DOI 10.1111/xen.12468.

Pera M.F., de Wert G., Dondorp W., Lovell-Badge R., Mummery C.L., Munsie M., Tam P.P. What if stem cells turn into embryos in a dish? Nat. Methods. 2015;12(10):917-919. DOI 10.1038/nmeth.3586.

Rashid T., Kobayashi T., Nakauchi H. Revisiting the flight of Icarus: making human organs from PSCs with large animal chimeras. Cell Stem Cell. 2014;15(4):406-409. DOI 10.1016/j.stem.2014.09.013.

Rivron N.C., Frias-Aldeguer J., Vrij E.J., Boisset J.C., Korving J., Vivié J., Truckenmüller R.K., van Oudenaarden A., van Blitterswijk C.A., Geijsen N. Blastocyst-like structures generated solely from stem cells. Nature. 2018;557(7703):106-111. DOI 10.1038/s41586-018-0051-0.

Shaw D., Dondorp W., Geijsen N., de Wert G. Creating human organs in chimaera pigs: an ethical source of immunocompatible organs? J. Med. Ethics. 2015;41(12):970-974. DOI 10.1136/medethics2014-102224.

Stanger B.Z., Tanaka A.J., Melton D.A. Organ size is limited by the number of embryonic progenitor cells in the pancreas but not the liver. Nature. 2007;445(7130):886-891. DOI 10.1038/nature05537.

Suchy F., Nakauchi H. Lessons from interspecies mammalian chimeras. Annu. Rev. Cell Dev. Biol. 2017;33:203-217. DOI 10.1146/annurevcellbio-100616-060654.

Sung Y.H., Baek I.J., Kim D.H., Jeon J., Lee J., Lee K., Jeong D., Kim J.S., Lee H.W. Knockout mice created by TALEN-mediated gene targeting. Nat. Biotechnol. 2013;31(1):23-24. DOI 10.1038/nbt.2477.

Tachibana M., Sparman M., Ramsey C., Ma H., Lee H.S., Penedo M.C., Mitalipov S. Generation of chimeric rhesus monkeys. Cell. 2012; 148(1-2):285-295. DOI 10.1016/j.cell.2011.12.007.

Takahashi K., Tanabe K., Ohnuki M., Narita M., Ichisaka T., Tomoda K., Yamanaka S. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell. 2007;131(5):861-872. DOI 10.1016/j.cell.2007.11.019.

Takahashi K., Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006;126(4):663-676. DOI 10.1016/j.cell.2006.07.024.

Tarkowski A.K. Mouse chimaeras developed from fused eggs. Nature. 1961;190:857-860. DOI 10.1038/190857a0.

Tesar P.J., Chenoweth J.G., Brook F.A., Davies T.J., Evans E.P., Mack D.L., Gardner R.L., McKay R.D. New cell lines from mouse epiblast share defining features with human embryonic stem cells. Nature. 2007;448(7150):196-199. DOI 10.1038/nature05972.

Theunissen T.W., Friedli M., He Y., Planet E., O’Neil R.C., Markoulaki S., Pontis J., Wang H., Iouranova A., Imbeault M., Duc J., Cohen M.A., Wert K.J., Castanon R., Zhang Z., Huang Y., Nery J.R., Drotar J., Lungjangwa T., Trono D., Ecker J.R., Jaenisch R. Molecular criteria for defining the naive human pluripotent state. Cell Stem Cell. 2016;19(4):502-515. DOI 10.1016/j.stem.2016.06.011.

Thomson J.A., Itskovitz-Eldor J., Shapiro S.S., Waknitz M.A., Swiergiel J.J., Marshall V.S., Jones J.M. Embryonic stem cell lines derived from human blastocysts. Science. 1998;282(5391):1145-1147. DOI 10.1126/science.282.5391.1145.

Thomson J.A., Kalishman J., Golos T.G., Durning M., Harris C.P., Becker R.A., Hearn J.P. Isolation of a primate embryonic stem cell line. Proc. Natl. Acad. Sci. USA. 1995;92(17):7844-7848. DOI 10.1073/pnas.92.17.7844.

Tippett P. Blood group chimeras. A review. Vox Sang. 1983;44(6):333- 359. DOI 10.1111/j.1423-0410.1983.tb03657.x.

Tsukiyama T., Ohinata Y. A modified EpiSC culture condition containing a GSK3 inhibitor can support germline-competent pluripotency in mice. PLoS One. 2014;9(4):e95329. DOI 10.1371/journal.pone.0095329.

Usui J., Kobayashi T., Yamaguchi T., Knisely A.S., Nishinakamura R., Nakauchi H. Generation of kidney from pluripotent stem cells via blastocyst complementation. Am. J. Pathol. 2012;180(6):2417- 2426. DOI 10.1016/j.ajpath.2012.03.007.

Wang R., Li T. DNA methylation is correlated with pluripotency of stem cells. Curr. Stem Cell Res. Ther. 2017;12(6):442-446. DOI 10.2174/1574888X11666161226145432.

Watanabe K., Ueno M., Kamiya D., Nishiyama A., Matsumura M., Wataya T., Takahashi J.B., Nishikawa S., Nishikawa S., Muguruma K., Sasai Y. A ROCK inhibitor permits survival of dissociated human embryonic stem cells. Nat. Biotechnol. 2007;25(6):681-686. DOI 10.1038/nbt1310.

Whitworth K.M., Lee K., Benne J.A., Beaton B.P., Spate L.D., Murphy S.L., Samuel M.S., Mao J., O’Gorman C., Walters E.M., Murphy C.N., Driver J., Mileham A., McLaren D., Wells K.D., Prather R.S. Use of the CRISPR/Cas9 system to produce genetically engineered pigs from in vitro-derived oocytes and embryos. Biol. Reprod. 2014;91(3):78. DOI 10.1095/biolreprod.114.121723.

Wobus A.M., Boheler K.R. Embryonic stem cells: prospects for developmental biology and cell therapy. Physiol. Rev. 2005;85(2):635- 678. DOI 10.1152/physrev.00054.2003.

Wu J., Platero-Luengo A., Sakurai M., Sugawara A., Gil M.A., Yamauchi T., Suzuki K., Bogliotti Y.S., Cuello C., Morales Valencia M., Okumura D., Luo J., Vilariño M., Parrilla I., Soto D.A., Martinez C.A., Hishida T., Sánchez-Bautista S., Martinez-Martinez M.L., Wang H., Nohalez A., Aizawa E., Martinez-Redondo P., Ocampo A., Reddy P., Roca J., Maga E.A., Esteban C.R., Berggren W.T., Nuñez Delicado E., Lajara J., Guillen I., Guillen P., Campistol J.M., Martinez E.A., Ross P.J., Izpisua Belmonte J.C. Interspecies chimerism with mammalian pluripotent stem cells. Cell. 2017;168(3):473-486. e415. DOI 10.1016/j.cell.2016.12.036.

Yamaguchi T., Sato H., Kato-Itoh M., Goto T., Hara H., Sanbo M., Mizuno N., Kobayashi T., Yanagida A., Umino A., Ota Y., Hamanaka S., Masaki H., Rashid S.T., Hirabayashi M., Nakauchi H. Interspecies organogenesis generates autologous functional islets. Nature. 2017;542(7640):191-196. DOI 10.1038/nature21070.

Yu J., Vodyanik M.A., Smuga-Otto K., Antosiewicz-Bourget J., Frane J.L., Tian S., Nie J., Jonsdottir G.A., Ruotti V., Stewart R., Slukvin I.I., Thomson J.A. Induced pluripotent stem cell lines derived from human somatic cells. Science. 2007;318(5858):1917-1920. DOI 10.1126/science.1151526.

Zhou W., Choi M., Margineantu D., Margaretha L., Hesson J., Cavanaugh C., Blau C.A., Horwitz M.S., Hockenbery D., Ware C., Ruohola-Baker H. HIF1α induced switch from bivalent to exclusively glycolytic metabolism during ESC-to-EpiSC/hESC transition. EMBO J. 2012;31(9):2103-2116. DOI 10.1038/emboj.2012.71.

The authors declare that there are no conflicts of interest present.