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

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 [∼43% of FACS-sorted embryoid body (EB) cells] from primed-state induced pluripotent stem cells (iPSCs) after a 72...

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Autores principales: Mitsunaga, Shino, Odajima, Junko, Yawata, Shiomi, Shioda, Keiko, Owa, Chie, Isselbacher, Kurt J., Hanna, Jacob H., Shioda, Toshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5699045/
https://www.ncbi.nlm.nih.gov/pubmed/29087313
http://dx.doi.org/10.1073/pnas.1707779114
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author Mitsunaga, Shino
Odajima, Junko
Yawata, Shiomi
Shioda, Keiko
Owa, Chie
Isselbacher, Kurt J.
Hanna, Jacob H.
Shioda, Toshi
author_facet Mitsunaga, Shino
Odajima, Junko
Yawata, Shiomi
Shioda, Keiko
Owa, Chie
Isselbacher, Kurt J.
Hanna, Jacob H.
Shioda, Toshi
author_sort Mitsunaga, Shino
collection PubMed
description 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 [∼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)-naïve 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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spelling pubmed-56990452017-11-27 Relevance of iPSC-derived human PGC-like cells at the surface of embryoid bodies to prechemotaxis migrating PGCs Mitsunaga, Shino Odajima, Junko Yawata, Shiomi Shioda, Keiko Owa, Chie Isselbacher, Kurt J. Hanna, Jacob H. Shioda, Toshi Proc Natl Acad Sci U S A PNAS Plus 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 [∼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)-naïve 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. National Academy of Sciences 2017-11-14 2017-10-30 /pmc/articles/PMC5699045/ /pubmed/29087313 http://dx.doi.org/10.1073/pnas.1707779114 Text en Copyright © 2017 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/ This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle PNAS Plus
Mitsunaga, Shino
Odajima, Junko
Yawata, Shiomi
Shioda, Keiko
Owa, Chie
Isselbacher, Kurt J.
Hanna, Jacob H.
Shioda, Toshi
Relevance of iPSC-derived human PGC-like cells at the surface of embryoid bodies to prechemotaxis migrating PGCs
title Relevance of iPSC-derived human PGC-like cells at the surface of embryoid bodies to prechemotaxis migrating PGCs
title_full Relevance of iPSC-derived human PGC-like cells at the surface of embryoid bodies to prechemotaxis migrating PGCs
title_fullStr Relevance of iPSC-derived human PGC-like cells at the surface of embryoid bodies to prechemotaxis migrating PGCs
title_full_unstemmed Relevance of iPSC-derived human PGC-like cells at the surface of embryoid bodies to prechemotaxis migrating PGCs
title_short Relevance of iPSC-derived human PGC-like cells at the surface of embryoid bodies to prechemotaxis migrating PGCs
title_sort relevance of ipsc-derived human pgc-like cells at the surface of embryoid bodies to prechemotaxis migrating pgcs
topic PNAS Plus
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5699045/
https://www.ncbi.nlm.nih.gov/pubmed/29087313
http://dx.doi.org/10.1073/pnas.1707779114
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