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Activin A-derived human embryonic stem cells show increased competence to differentiate into primordial germ cell-like cells

Activin A-derived human embryonic stem cells show increased competence to differentiate into primordial germ cell-like cells

Authors :
Filip Van Nieuwerburgh
Annekatrien Boel
Susana M. Chuva de Sousa Lopes
Dieter Deforce
Margot Van der Jeught
Laurentijn Tilleman
Jasin Taelman
Petra De Sutter
Swati Mishra
Mina Popovic
Björn Menten
Evi Duthoo
Björn Heindryckx
Source :
STEM CELLS, Stem Cells (Dayton, Ohio), Stem Cells, 39(5), 551-563. OXFORD UNIVERSITY PRESS
Publication Year :
2021
Publisher :
OXFORD UNIVERSITY PRESS, 2021.

Abstract

Protocols for specifying human primordial germ cell‐like cells (hPGCLCs) from human embryonic stem cells (hESCs) remain hindered by differences between hESC lines, their derivation methods, and maintenance culture conditions. This poses significant challenges for establishing reproducible in vitro models of human gametogenesis. Here, we investigated the influence of activin A (ActA) during derivation and maintenance on the propensity of hESCs to differentiate into PGCLCs. We show that continuous ActA supplementation during hESC derivation (from blastocyst until the formation of the post‐inner cell mass intermediate [PICMI]) and supplementation (from the first passage of the PICMI onwards) is beneficial to differentiate hESCs to PGCLCs subsequently. Moreover, comparing isogenic primed and naïve states prior to differentiation, we showed that conversion of hESCs to the 4i‐state improves differentiation to (TNAP [tissue nonspecific alkaline phosphatase]+/PDPN [podoplanin]+) PGCLCs. Those PGCLCs expressed several germ cell markers, including TFAP2C (transcription factor AP‐2 gamma), SOX17 (SRY‐box transcription factor 17), and NANOS3 (nanos C2HC‐type zinc finger 3), and markers associated with germ cell migration, CXCR4 (C‐X‐C motif chemokine receptor 4), LAMA4 (laminin subunit alpha 4), ITGA6 (integrin subunit alpha 6), and CDH4 (cadherin 4), suggesting that the large numbers of PGCLCs obtained may be suitable to differentiate further into more mature germ cells. Finally, hESCs derived in the presence of ActA showed higher competence to differentiate to hPGCLC, in particular if transiently converted to the 4i‐state. Our work provides insights into the differences in differentiation propensity of hESCs and delivers an optimized protocol to support efficient human germ cell derivation.<br />Human embryonic stem cells (hESCs) derived and cultured in the presence of activin A show higher propensity to differentiate to (TNAP+PDPN+) human primordial germ cell‐like cells, particularly when transiently converted to the 4i‐state prior to differentiation using an embryoid body aggregation assay.

Details

Language :
English
ISSN :
10665099 and 15494918
Database :
OpenAIRE
Journal :
STEM CELLS, Stem Cells (Dayton, Ohio), Stem Cells, 39(5), 551-563. OXFORD UNIVERSITY PRESS
Accession number :
edsair.doi.dedup.....0fd0d34e28914dfd8f37facfd4a5a769