Relevance of iPSC-derived human PGC-like cells at the surface of embryoid bodies to prechemotaxis migrating PGCs

Mitsunaga Shino; Odajima Junko; Yawata ShiomiShioda KeikoOwa ChieIsselbacher Kurt J.Hanna Jacob H.Shioda Toshi

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Relevance of iPSC-derived human PGC-like cells at the surface of embryoid bodies to prechemotaxis migrating PGCs

机译：Relevance of iPSC-derived human PGC-like cells at the surface of embryoid bodies to prechemotaxis migrating PGCs

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Pluripotent stem cell-derived human primordial germ cell-like cells (hPGCLCs) provide important opportunities to study primordial germ cells (PGCs). We robustly produced CD38(+) hPGCLCs similar to 43 of FACS-sorted embryoid body (EB) cells from primed-state induced pluripotent stem cells (iPSCs) after a 72-hour transient incubation in the four chemical inhibitors (4i)-naive reprogramming medium and showed transcriptional consistency of our hPGCLCs with hPGCLCs generated in previous studies using various and distinct protocols. Both CD38(+) hPGCLCs and CD38(-) EB cells significantly expressed PRDM1 and TFAP2C, although PRDM1 mRNA in CD38(-) cells lacked the 3'-UTR harboring miRNA binding sites regulating mRNA stability. Genes up-regulated in hPGCLCs were enriched for cell migration genes, and their promoters were enriched for the binding motifs of TFAP2 (which was identified in promoters of T, NANOS3, and SOX17) and the RREB-1 cell adhesion regulator. In EBs, hPGCLCs were identified exclusively in the outermost surface monolayer as dispersed cells or cell aggregates with strong and specific expression of POU5F1/OCT4 protein. Time-lapse live cell imaging revealed active migration of hPGCLCs on Matrigel. Whereas all hPGCLCs strongly expressed the CXCR4 chemotaxis receptor, its ligand CXCL12/SDF1 was not significantly expressed in the whole EBs. Exposure of hPGCLCs to CXCL12/SDF1 induced cell migration genes and antiapoptosis genes. Thus, our study shows that transcriptionally consistent hPGCLCs can be readily produced from hiPSCs after transition of their pluripotency from the primed state using various methods and that hPGCLCs resemble the early-stage PGCs randomly migrating in the midline region of human embryos before initiation of the CXCL12/SDF1-guided chemotaxis.

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《Proceedings of the National Academy of Sciences of the United States of America.》 |2017年第46期|E9913-E9922|共10页
作者
Mitsunaga Shino; Odajima Junko; Yawata ShiomiShioda KeikoOwa ChieIsselbacher Kurt J.Hanna Jacob H.Shioda Toshi;
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作者单位

Weizmann Inst Sci, Dept Mol Genet, IL-76100 Rehovot, Israel;

Massachusetts Gen Hosp, Ctr Canc Res, Charlestown, MA 02129 USA;

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收录信息美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
原文格式 PDF
正文语种英语
中图分类自然科学总论;
关键词
primordial germ cells; induced pluripotent stem cells; embryoid body; PRDM1; TFAP2C;

Relevance of iPSC-derived human PGC-like cells at the surface of embryoid bodies to prechemotaxis migrating PGCs

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