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Induction of mouse germ-cell fate by transcription factors in vitro


The germ-cell lineage ensures the continuity of life through the generation of male and female gametes, which unite to form a totipotent zygote. We have previously demonstrated that, by using cytokines, embryonic stem cells and induced pluripotent stem cells can be induced into epiblast-like cells (EpiLCs) and then into primordial germ cell (PGC)-like cells with the capacity for both spermatogenesis and oogenesis1,2, creating an opportunity for understanding and regulating mammalian germ-cell development in both sexes in vitro. Here we show that, without cytokines, simultaneous overexpression of three transcription factors, Blimp1 (also known as Prdm1), Prdm14 and Tfap2c (also known as AP2γ), directs EpiLCs, but not embryonic stem cells, swiftly and efficiently into a PGC state. Notably, Prdm14 alone, but not Blimp1 or Tfap2c, suffices for the induction of the PGC state in EpiLCs. The transcription-factor-induced PGC state, irrespective of the transcription factors used, reconstitutes key transcriptome and epigenetic reprogramming in PGCs, but bypasses a mesodermal program that accompanies PGC or PGC-like-cell specification by cytokines including bone morphogenetic protein 4. Notably, the transcription-factor-induced PGC-like cells contribute to spermatogenesis and fertile offspring. Our findings provide a new insight into the transcriptional logic for PGC specification, and create a foundation for the transcription-factor-based reconstitution and regulation of mammalian gametogenesis.

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Figure 1: Induction of a PGC-like state by TFs.
Figure 2: Global transcription profiles for TF- and Ck-PGCLCs and epigenetic properties of TF-PGCLCs.
Figure 3: Global transcriptional target analyses of the three TFs.
Figure 4: Spermatogenesis and fertile offspring from TF (BP14A)-PGCLCs.

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Gene Expression Omnibus

Referenced accessions

Gene Expression Omnibus

Data deposits

The accession number for the microarray data presented in this study is available fromthe Gene ExpressionOmnibus (GEO) database under accession GSE46855.


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We thank A. Bradley, A. Smith, G. Guo, H. Niwa and G. Nagamatsu for providing plasmids. We are grateful to the Center for Anatomical Studies (Graduate School of Medicine, Kyoto University) for performing the histological analyses. We thank M. Yamaji for advice and T. Mori for encouragement. F.N. is a Japan Society for the Promotion of Science (JSPS) Research Fellow. This study was supported in part by a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan; by JST-CREST/ERATO; by the Takeda Science Foundation; and by the Academia for Repro-regenerative Medicine.

Author information

Authors and Affiliations



F.N. designed and conducted the experiments, analysed the data, and wrote the manuscript. K.H. designed the experiments and analysed the data. K.K. conducted the microarray experiments. H.O. performed the transplantation of cells into seminiferous tubules. Y.Y. analysed the data. M.S. conceived the project, designed the experiments and wrote the manuscript.

Corresponding author

Correspondence to Mitinori Saitou.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

This file contains Supplementary Figures 1-10. (PDF 5684 kb)

Supplementary Table 1

This file contains the top 100 genes contributing to the PC1 and PC2 of Figure 2a. (XLS 51 kb)

Supplementary Table 2

This file contains core PGC genes and somatic mesodermal genes (related to Supplementary Figure 6b). (XLS 78 kb)

Supplementary Table 3

This file contains the top 100 genes up-/down-regulated in TF(BP14A)-PGCLCs compared to EpiLCs and in SC-ESCs by BP14A compared to ESCs (related to Supplementary Figure 6c). (XLS 74 kb)

Supplementary Table 4

This file contains the top 100 genes contributing to the PC1 and PC2 of Figure 3a. (XLS 50 kb)

Supplementary Table 5

This file contains lists of genes exhibiting differential expression between P14 and B cells (related to Supplementary Figure 9). (XLS 102 kb)

Supplementary Table 6

This file contains spermatogenesis by TF- or Ck-PGCLCs. (XLS 22 kb)

Supplementary Table 7

This file contains development of embryos derived from TF- and Ck-PGCLC-derived spermatozoa. (XLS 20 kb)

Supplementary Table 8

This file contains tagged sequences for exogenous transcription factors. (XLS 14 kb)

Supplementary Table 9

This file contains primer sequences used in this study. (XLS 24 kb)

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Nakaki, F., Hayashi, K., Ohta, H. et al. Induction of mouse germ-cell fate by transcription factors in vitro . Nature 501, 222–226 (2013).

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