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

Proceedings of the National Academy of Sciences of the United States of America
Shino MitsunagaToshi Shioda

Abstract

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 th...Continue Reading

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Citations

Sep 19, 2018·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·Haihan Tan, Wee-Wei Tee
Mar 3, 2020·Tissue Engineering and Regenerative Medicine·Marina V Pryzhkova, Philip W Jordan
Jul 28, 2019·International Journal of Molecular Sciences·Zhenwu ZhangChao-Po Lin
Jul 10, 2018·Nucleic Acids Research·Yishai YehudaItamar Simon
Mar 22, 2019·Human Reproduction Open·C EguizabalA Veiga
Jul 17, 2020·Frontiers in Bioengineering and Biotechnology·Sneha GopalJonathan S Dordick
May 19, 2020·The Journal of Reproduction and Development·Maria SorrentiMassimo DE Felici
Jun 1, 2021·Frontiers in Cell and Developmental Biology·Lucia A Torres-FernándezWaldemar Kolanus
Dec 3, 2021·Development·Christopher B Cooke, Naomi Moris

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Methods Mentioned

BETA
FACS
PCA
RNA-seq
fluorescence imaging

Software Mentioned

edgeR
R
STAR aligner
Rsubread
Bioconductor
DAVID
plot3D
prcomp
TRANSFAC

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