Differentiation of Human-Induced Pluripotent Stem Cells (hiPSCs) into Human Primordial Germ Cell-like Cells (hPGCLCs) In Vitro

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2022 Apr 29

PMID 35486250

Authors

Ahmed Reda

Jan-Bernd Stukenborg

Qiaolin Deng

Affiliations

Soon will be listed here.

Abstract

In humans, germ cells are specified in the extraembryonic yolk sac, at proximity of allantois, around the second week of gestation. Derivation of human germ cell-like cells (hPGCLCs) from human pluripotent cells in vitro is of a great importance for research purposes, such as disease modeling, or studying the early human germ cell development and the effect of environmental factors on this development. As it is not possible to access human embryos at early developmental stages, a two-step protocol has been proposed by Sasaki and colleagues to differentiate hPGCLCs in vitro from human pluripotent stem cells. Here, we report a detailed protocol for in vitro hPGCLCs differentiation from induced pluripotent stem cells (iPSCs).

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References

Tang W, Kobayashi T, Irie N, Dietmann S, Surani M . Specification and epigenetic programming of the human germ line. Nat Rev Genet. 2016; 17(10):585-600. DOI: 10.1038/nrg.2016.88. View

Leitch H, Tang W, Surani M . Primordial germ-cell development and epigenetic reprogramming in mammals. Curr Top Dev Biol. 2013; 104:149-87. DOI: 10.1016/B978-0-12-416027-9.00005-X. View

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. DOI: 10.1016/j.cell.2009.03.014. View

Ying Y, Zhao G . Cooperation of endoderm-derived BMP2 and extraembryonic ectoderm-derived BMP4 in primordial germ cell generation in the mouse. Dev Biol. 2001; 232(2):484-92. DOI: 10.1006/dbio.2001.0173. View

Lawson K, Dunn N, Roelen B, Zeinstra L, Davis A, Wright C . Bmp4 is required for the generation of primordial germ cells in the mouse embryo. Genes Dev. 1999; 13(4):424-36. PMC: 316469. DOI: 10.1101/gad.13.4.424. View

Ying Y, Liu X, Marble A, Lawson K, Zhao G . Requirement of Bmp8b for the generation of primordial germ cells in the mouse. Mol Endocrinol. 2000; 14(7):1053-63. DOI: 10.1210/mend.14.7.0479. View

Ohinata Y, Payer B, OCarroll D, Ancelin K, Ono Y, Sano M . Blimp1 is a critical determinant of the germ cell lineage in mice. Nature. 2005; 436(7048):207-13. DOI: 10.1038/nature03813. View

Magnusdottir E, Dietmann S, Murakami K, Gunesdogan U, Tang F, Bao S . A tripartite transcription factor network regulates primordial germ cell specification in mice. Nat Cell Biol. 2013; 15(8):905-15. PMC: 3796875. DOI: 10.1038/ncb2798. View

Irie N, Weinberger L, Tang W, Kobayashi T, Viukov S, Manor Y . SOX17 is a critical specifier of human primordial germ cell fate. Cell. 2014; 160(1-2):253-68. PMC: 4310934. DOI: 10.1016/j.cell.2014.12.013. View

10.

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. DOI: 10.1016/j.cell.2011.06.052. View

11.

Kee K, Angeles V, Flores M, Nguyen H, Pera R . Human DAZL, DAZ and BOULE genes modulate primordial germ-cell and haploid gamete formation. Nature. 2009; 462(7270):222-5. PMC: 3133736. DOI: 10.1038/nature08562. View

12.

Gafni O, Weinberger L, AlFatah Mansour A, Manor Y, Chomsky E, Ben-Yosef D . Derivation of novel human ground state naive pluripotent stem cells. Nature. 2013; 504(7479):282-6. DOI: 10.1038/nature12745. View

13.

Sasaki K, Yokobayashi S, Nakamura T, Okamoto I, Yabuta Y, Kurimoto K . Robust In Vitro Induction of Human Germ Cell Fate from Pluripotent Stem Cells. Cell Stem Cell. 2015; 17(2):178-94. DOI: 10.1016/j.stem.2015.06.014. View

14.

Sugawa F, Arauzo-Bravo M, Yoon J, Kim K, Aramaki S, Wu G . Human primordial germ cell commitment in vitro associates with a unique PRDM14 expression profile. EMBO J. 2015; 34(8):1009-24. PMC: 4406649. DOI: 10.15252/embj.201488049. View

15.

Chen D, Liu W, Lukianchikov A, Hancock G, Zimmerman J, Lowe M . Germline competency of human embryonic stem cells depends on eomesodermin. Biol Reprod. 2017; 97(6):850-861. PMC: 5803789. DOI: 10.1093/biolre/iox138. View

16.

Panula S, Kurek M, Kumar P, Albalushi H, Sanchez S, Damdimopoulou P . Human induced pluripotent stem cells from two azoospermic patients with Klinefelter syndrome show similar X chromosome inactivation behavior to female pluripotent stem cells. Hum Reprod. 2019; 34(11):2297-2310. PMC: 6894010. DOI: 10.1093/humrep/dez134. View

17.

Okita K, Matsumura Y, Sato Y, Okada A, Morizane A, Okamoto S . A more efficient method to generate integration-free human iPS cells. Nat Methods. 2011; 8(5):409-12. DOI: 10.1038/nmeth.1591. View

18.

Ishizaki T, Uehata M, Tamechika I, Keel J, Nonomura K, Maekawa M . Pharmacological properties of Y-27632, a specific inhibitor of rho-associated kinases. Mol Pharmacol. 2000; 57(5):976-83. View

19.

DAVIES S, Reddy H, Caivano M, Cohen P . Specificity and mechanism of action of some commonly used protein kinase inhibitors. Biochem J. 2000; 351(Pt 1):95-105. PMC: 1221339. DOI: 10.1042/0264-6021:3510095. View