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SOX17 is a critical specifier of human primordial germ cell fate.

Irie N, Weinberger L, Tang WW, Kobayashi T, Viukov S, Manor YS, Dietmann S, Hanna JH, Surani MA - Cell (2014)

Bottom Line: Here, we demonstrate specification of hPGC-like cells (hPGCLCs) from germline competent pluripotent stem cells.Remarkably, SOX17 is the key regulator of hPGC-like fate, whereas BLIMP1 represses endodermal and other somatic genes during specification of hPGCLCs.We have established a foundation for future studies on resetting of the epigenome in hPGCLCs and hPGCs for totipotency and the transmission of genetic and epigenetic information.

View Article: PubMed Central - PubMed

Affiliation: Wellcome Trust Cancer Research UK Gurdon Institute, Tennis Court Road, University of Cambridge, Cambridge CB2 1QN, UK; Department of Physiology, Development and Neuroscience, Downing Street, University of Cambridge, Cambridge CB2 3EG, UK; Wellcome Trust-Medical Research Council Stem Cell Institute, Tennis Court Road, University of Cambridge, Cambridge CB2 3EG, UK.

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Sequential Expression of Germ-Cell-Related Transcription Factors in Single Cells during hPGCLC Specification(A and B) Immunofluorscence analysis for (A) BLIMP1, SOX17, and TFAP2C and (B) BLIMP1 and T in cryosections of day 1–8 embryoids after hPGCLC induction. Bottom row in (B) shows high exposure (digital) image of T, indicating low but specific expression in hPGCLC. SOX17-positive or BLIMP1-positive cells are highlighted. Scale bars, 50 μm.(C) Percentage of SOX17-positive (+) cells in day 1–8 embryoids that were also TFAP2C+ or BLIMP1+. Corresponds to data in Figure 4A.(D) Percentage of BLIMP1-positive (+) cells in day 1–8 embryoids that were TFAP2C+, NANOG+, or OCT4+. Corresponds to data in Figures 4A, S4A, and S4B.(E) Summary model for dynamics of hPGCLC specification in embryoids. SOX17-positive cells are first scattered in day 1 embryoids. They gain expression of BLIMP1, TFAP2C, and NANOG sequentially and form a cluster from day 2 onward until the formation of nascent hPGCLC.
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fig4: Sequential Expression of Germ-Cell-Related Transcription Factors in Single Cells during hPGCLC Specification(A and B) Immunofluorscence analysis for (A) BLIMP1, SOX17, and TFAP2C and (B) BLIMP1 and T in cryosections of day 1–8 embryoids after hPGCLC induction. Bottom row in (B) shows high exposure (digital) image of T, indicating low but specific expression in hPGCLC. SOX17-positive or BLIMP1-positive cells are highlighted. Scale bars, 50 μm.(C) Percentage of SOX17-positive (+) cells in day 1–8 embryoids that were also TFAP2C+ or BLIMP1+. Corresponds to data in Figure 4A.(D) Percentage of BLIMP1-positive (+) cells in day 1–8 embryoids that were TFAP2C+, NANOG+, or OCT4+. Corresponds to data in Figures 4A, S4A, and S4B.(E) Summary model for dynamics of hPGCLC specification in embryoids. SOX17-positive cells are first scattered in day 1 embryoids. They gain expression of BLIMP1, TFAP2C, and NANOG sequentially and form a cluster from day 2 onward until the formation of nascent hPGCLC.

Mentions: On day 1, we first detected SOX17 in a few widely scattered cells throughout the embryoids (Figures 4A and 4E). Among the SOX17-positive (+) cells, 55% were also BLIMP1+, and 22% were TFAP2C+ (Figures 4A and 4C). However, all BLIMP1+ cells coexpressed SOX17, suggesting that SOX17 is upregulated before BLIMP1. The proportion of BLIMP1+ and TFAP2C+ cells increased to ∼70% on day 2 and to ∼90% on days 4–8 (Figures 4A and 4C). These triple-positive cells likely represent specified hPGCLCs, as they also coexpressed other key hPGC genes. However, ∼10% of single SOX17+ cells failed to undergo hPGCLC specification but persisted in day 4–8 embryoids. These may be aberrant cells or else may belong to other lineages.


SOX17 is a critical specifier of human primordial germ cell fate.

Irie N, Weinberger L, Tang WW, Kobayashi T, Viukov S, Manor YS, Dietmann S, Hanna JH, Surani MA - Cell (2014)

Sequential Expression of Germ-Cell-Related Transcription Factors in Single Cells during hPGCLC Specification(A and B) Immunofluorscence analysis for (A) BLIMP1, SOX17, and TFAP2C and (B) BLIMP1 and T in cryosections of day 1–8 embryoids after hPGCLC induction. Bottom row in (B) shows high exposure (digital) image of T, indicating low but specific expression in hPGCLC. SOX17-positive or BLIMP1-positive cells are highlighted. Scale bars, 50 μm.(C) Percentage of SOX17-positive (+) cells in day 1–8 embryoids that were also TFAP2C+ or BLIMP1+. Corresponds to data in Figure 4A.(D) Percentage of BLIMP1-positive (+) cells in day 1–8 embryoids that were TFAP2C+, NANOG+, or OCT4+. Corresponds to data in Figures 4A, S4A, and S4B.(E) Summary model for dynamics of hPGCLC specification in embryoids. SOX17-positive cells are first scattered in day 1 embryoids. They gain expression of BLIMP1, TFAP2C, and NANOG sequentially and form a cluster from day 2 onward until the formation of nascent hPGCLC.
© Copyright Policy - CC BY
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4310934&req=5

fig4: Sequential Expression of Germ-Cell-Related Transcription Factors in Single Cells during hPGCLC Specification(A and B) Immunofluorscence analysis for (A) BLIMP1, SOX17, and TFAP2C and (B) BLIMP1 and T in cryosections of day 1–8 embryoids after hPGCLC induction. Bottom row in (B) shows high exposure (digital) image of T, indicating low but specific expression in hPGCLC. SOX17-positive or BLIMP1-positive cells are highlighted. Scale bars, 50 μm.(C) Percentage of SOX17-positive (+) cells in day 1–8 embryoids that were also TFAP2C+ or BLIMP1+. Corresponds to data in Figure 4A.(D) Percentage of BLIMP1-positive (+) cells in day 1–8 embryoids that were TFAP2C+, NANOG+, or OCT4+. Corresponds to data in Figures 4A, S4A, and S4B.(E) Summary model for dynamics of hPGCLC specification in embryoids. SOX17-positive cells are first scattered in day 1 embryoids. They gain expression of BLIMP1, TFAP2C, and NANOG sequentially and form a cluster from day 2 onward until the formation of nascent hPGCLC.
Mentions: On day 1, we first detected SOX17 in a few widely scattered cells throughout the embryoids (Figures 4A and 4E). Among the SOX17-positive (+) cells, 55% were also BLIMP1+, and 22% were TFAP2C+ (Figures 4A and 4C). However, all BLIMP1+ cells coexpressed SOX17, suggesting that SOX17 is upregulated before BLIMP1. The proportion of BLIMP1+ and TFAP2C+ cells increased to ∼70% on day 2 and to ∼90% on days 4–8 (Figures 4A and 4C). These triple-positive cells likely represent specified hPGCLCs, as they also coexpressed other key hPGC genes. However, ∼10% of single SOX17+ cells failed to undergo hPGCLC specification but persisted in day 4–8 embryoids. These may be aberrant cells or else may belong to other lineages.

Bottom Line: Here, we demonstrate specification of hPGC-like cells (hPGCLCs) from germline competent pluripotent stem cells.Remarkably, SOX17 is the key regulator of hPGC-like fate, whereas BLIMP1 represses endodermal and other somatic genes during specification of hPGCLCs.We have established a foundation for future studies on resetting of the epigenome in hPGCLCs and hPGCs for totipotency and the transmission of genetic and epigenetic information.

View Article: PubMed Central - PubMed

Affiliation: Wellcome Trust Cancer Research UK Gurdon Institute, Tennis Court Road, University of Cambridge, Cambridge CB2 1QN, UK; Department of Physiology, Development and Neuroscience, Downing Street, University of Cambridge, Cambridge CB2 3EG, UK; Wellcome Trust-Medical Research Council Stem Cell Institute, Tennis Court Road, University of Cambridge, Cambridge CB2 3EG, UK.

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Related in: MedlinePlus