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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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Comparison of hESC Transcriptomes under Conventional or 4i Culture Conditions, Related to Figure 7 and Table S2(A) Scatter plot showing global gene expression levels (mean log2(normalized read counts) of two replicates) between WIS2 hESC cultured under 4i and conventional condition. Each dot represents one gene. Differentially expressed genes (log2(fold change) > 2 and adjusted p value < 0.05) were presented as red dots (upregulated in 4i condition) or blue dots (upregulated in conventional condition).(B) Gene ontology (GO) term enrichment analysis of upregulated genes in 4i WIS2 hESC (upper panel) and conventional WIS2 hESC (lower panel). Top 15 GO biological process terms that were enriched in each condition were shown (DAVID GOTERM_BP_FAT with gene count > = 5 followed by GO Trimming to reduce term redundancy).(C) Heat map showing expression of representative pluripotency and mesodermal genes expression in WIS2 hESCs cultured under 4i or conventional conditions (Conv). RNASeq data from two biological replicates were shown (#1 and #2). Asterisk indicates differential expression with statistical significance (log2(fold change) > 2 and adjusted p value < 0.05).(D) Immunofluorescence analysis of T and OCT4 on WIS2 hESCs cultured under conventional (Conv hESC) and 4i condition (4i hESC).
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figs7: Comparison of hESC Transcriptomes under Conventional or 4i Culture Conditions, Related to Figure 7 and Table S2(A) Scatter plot showing global gene expression levels (mean log2(normalized read counts) of two replicates) between WIS2 hESC cultured under 4i and conventional condition. Each dot represents one gene. Differentially expressed genes (log2(fold change) > 2 and adjusted p value < 0.05) were presented as red dots (upregulated in 4i condition) or blue dots (upregulated in conventional condition).(B) Gene ontology (GO) term enrichment analysis of upregulated genes in 4i WIS2 hESC (upper panel) and conventional WIS2 hESC (lower panel). Top 15 GO biological process terms that were enriched in each condition were shown (DAVID GOTERM_BP_FAT with gene count > = 5 followed by GO Trimming to reduce term redundancy).(C) Heat map showing expression of representative pluripotency and mesodermal genes expression in WIS2 hESCs cultured under 4i or conventional conditions (Conv). RNASeq data from two biological replicates were shown (#1 and #2). Asterisk indicates differential expression with statistical significance (log2(fold change) > 2 and adjusted p value < 0.05).(D) Immunofluorescence analysis of T and OCT4 on WIS2 hESCs cultured under conventional (Conv hESC) and 4i condition (4i hESC).

Mentions: Global gene expression analysis indicated overall similarities between hESCs in the conventional medium versus those in “4i” medium (r = 0.923) but with notable differences (Figure S7A). Although these cells showed similar expression levels of core pluripotency factors OCT4, NANOG, and SOX2, 4i hESCs had higher expression of mesoderm and gastrulation genes, including T, RUNX1, and PDGFRA (Figures S7B and S7C and Table S2). Furthermore, OCT4-positive cells in 4i hESCs had varying levels of T protein, possibly due to inhibition of GSK3β (Chen et al., 2013), which is not the case in hESC cultured in conventional condition (Figure S7D). These differences might be relevant for the mechanism of competence of ESCs for PGCLC, which merits further investigation.


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)

Comparison of hESC Transcriptomes under Conventional or 4i Culture Conditions, Related to Figure 7 and Table S2(A) Scatter plot showing global gene expression levels (mean log2(normalized read counts) of two replicates) between WIS2 hESC cultured under 4i and conventional condition. Each dot represents one gene. Differentially expressed genes (log2(fold change) > 2 and adjusted p value < 0.05) were presented as red dots (upregulated in 4i condition) or blue dots (upregulated in conventional condition).(B) Gene ontology (GO) term enrichment analysis of upregulated genes in 4i WIS2 hESC (upper panel) and conventional WIS2 hESC (lower panel). Top 15 GO biological process terms that were enriched in each condition were shown (DAVID GOTERM_BP_FAT with gene count > = 5 followed by GO Trimming to reduce term redundancy).(C) Heat map showing expression of representative pluripotency and mesodermal genes expression in WIS2 hESCs cultured under 4i or conventional conditions (Conv). RNASeq data from two biological replicates were shown (#1 and #2). Asterisk indicates differential expression with statistical significance (log2(fold change) > 2 and adjusted p value < 0.05).(D) Immunofluorescence analysis of T and OCT4 on WIS2 hESCs cultured under conventional (Conv hESC) and 4i condition (4i hESC).
© Copyright Policy - CC BY
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4310934&req=5

figs7: Comparison of hESC Transcriptomes under Conventional or 4i Culture Conditions, Related to Figure 7 and Table S2(A) Scatter plot showing global gene expression levels (mean log2(normalized read counts) of two replicates) between WIS2 hESC cultured under 4i and conventional condition. Each dot represents one gene. Differentially expressed genes (log2(fold change) > 2 and adjusted p value < 0.05) were presented as red dots (upregulated in 4i condition) or blue dots (upregulated in conventional condition).(B) Gene ontology (GO) term enrichment analysis of upregulated genes in 4i WIS2 hESC (upper panel) and conventional WIS2 hESC (lower panel). Top 15 GO biological process terms that were enriched in each condition were shown (DAVID GOTERM_BP_FAT with gene count > = 5 followed by GO Trimming to reduce term redundancy).(C) Heat map showing expression of representative pluripotency and mesodermal genes expression in WIS2 hESCs cultured under 4i or conventional conditions (Conv). RNASeq data from two biological replicates were shown (#1 and #2). Asterisk indicates differential expression with statistical significance (log2(fold change) > 2 and adjusted p value < 0.05).(D) Immunofluorescence analysis of T and OCT4 on WIS2 hESCs cultured under conventional (Conv hESC) and 4i condition (4i hESC).
Mentions: Global gene expression analysis indicated overall similarities between hESCs in the conventional medium versus those in “4i” medium (r = 0.923) but with notable differences (Figure S7A). Although these cells showed similar expression levels of core pluripotency factors OCT4, NANOG, and SOX2, 4i hESCs had higher expression of mesoderm and gastrulation genes, including T, RUNX1, and PDGFRA (Figures S7B and S7C and Table S2). Furthermore, OCT4-positive cells in 4i hESCs had varying levels of T protein, possibly due to inhibition of GSK3β (Chen et al., 2013), which is not the case in hESC cultured in conventional condition (Figure S7D). These differences might be relevant for the mechanism of competence of ESCs for PGCLC, which merits further investigation.

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