Activin A-derived human embryonic stem cells show increased competence to differentiate into primordial germ cell-like cells.
Activins
/ genetics
Blastocyst
/ cytology
Cadherins
/ genetics
Cell Differentiation
/ genetics
Cells, Cultured
Gene Expression Regulation, Developmental
/ genetics
Germ Cells
/ cytology
Human Embryonic Stem Cells
/ cytology
Humans
Integrin alpha6
/ genetics
Laminin
/ genetics
RNA-Binding Proteins
/ genetics
Receptors, CXCR4
/ genetics
SOXF Transcription Factors
/ genetics
Signal Transduction
/ genetics
Transcription Factor AP-2
/ genetics
activin A (ActA)
differentiation
human embryonic stem cells (hESCs)
pluripotency
primordial germ cell-like cells (PGCLCs)
Journal
Stem cells (Dayton, Ohio)
ISSN: 1549-4918
Titre abrégé: Stem Cells
Pays: England
ID NLM: 9304532
Informations de publication
Date de publication:
05 2021
05 2021
Historique:
received:
03
10
2020
accepted:
21
12
2020
pubmed:
21
1
2021
medline:
15
12
2021
entrez:
20
1
2021
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 33470497
doi: 10.1002/stem.3335
pmc: PMC8248136
doi:
Substances chimiques
CXCR4 protein, human
0
Cadherins
0
ITGA6 protein, human
0
Integrin alpha6
0
LAMA4 protein, human
0
Laminin
0
NANOS3 protein, human
0
R-cadherin
0
RNA-Binding Proteins
0
Receptors, CXCR4
0
SOX17 protein, human
0
SOXF Transcription Factors
0
TFAP2C protein, human
0
Transcription Factor AP-2
0
activin A
0
Activins
104625-48-1
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
551-563Informations de copyright
©2021 The Authors. Stem Cells published by Wiley Periodicals LLC on behalf of AlphaMed Press 2021.
Références
Nature. 2007 Jul 12;448(7150):191-5
pubmed: 17597762
Dev Biol. 1994 Aug;164(2):361-73
pubmed: 8045339
Oncogene. 2001 Jun 7;20(26):3332-40
pubmed: 11423983
Cell Stem Cell. 2016 Mar 3;18(3):330-40
pubmed: 26923202
Nat Methods. 2012 Jun 28;9(7):676-82
pubmed: 22743772
Cell Stem Cell. 2016 Jul 7;19(1):66-80
pubmed: 27320042
Development. 2017 Feb 1;144(3):365-373
pubmed: 28143843
Proc Natl Acad Sci U S A. 2017 Nov 14;114(46):E9913-E9922
pubmed: 29087313
Nature. 2016 Nov 10;539(7628):299-303
pubmed: 27750280
Cell Rep. 2019 Dec 24;29(13):4568-4582.e5
pubmed: 31875561
Bioessays. 2018 May;40(5):e1700239
pubmed: 29574793
Nature. 2013 Dec 12;504(7479):282-6
pubmed: 24172903
Mol Hum Reprod. 2018 Apr 1;24(4):173-184
pubmed: 29471503
Stem Cells Dev. 2019 May 1;28(9):579-592
pubmed: 30827199
Cell. 2015 Jan 15;160(1-2):253-68
pubmed: 25543152
Cell Stem Cell. 2017 Oct 5;21(4):517-532.e5
pubmed: 28985527
Cell Stem Cell. 2007 Sep 13;1(3):299-312
pubmed: 18371364
EMBO J. 2017 Jul 3;36(13):1888-1907
pubmed: 28559416
Hum Reprod Update. 2020 Sep 1;26(5):670-688
pubmed: 32464645
Science. 1998 Nov 6;282(5391):1145-7
pubmed: 9804556
Nat Commun. 2017 Oct 18;8(1):1034
pubmed: 29044119
Nat Rev Genet. 2016 Oct;17(10):585-600
pubmed: 27573372
Cell. 2011 Aug 19;146(4):519-32
pubmed: 21820164
Genes Cells. 2001 Nov;6(11):923-30
pubmed: 11733030
Stem Cell Reports. 2016 Apr 12;6(4):437-446
pubmed: 26947977
Development. 2015 Feb 15;142(4):607-19
pubmed: 25670788
Nucleic Acids Res. 2019 Feb 28;47(4):1605-1614
pubmed: 30566647
Science. 2012 Nov 16;338(6109):971-5
pubmed: 23042295
EMBO J. 1989 Sep;8(9):2543-50
pubmed: 2573523
Mol Cell Biol. 2004 May;24(10):4241-54
pubmed: 15121845
Nature. 2007 Jul 12;448(7150):196-9
pubmed: 17597760
Oncogene. 2003 Nov 6;22(50):8212-20
pubmed: 14603262
Nat Commun. 2020 Feb 7;11(1):764
pubmed: 32034154
EMBO J. 2015 Apr 15;34(8):1009-24
pubmed: 25750208
Cancers (Basel). 2018 Mar 28;10(4):
pubmed: 29597249
Stem Cell Reports. 2015 Jan 13;4(1):114-128
pubmed: 25544567
Nature. 1998 Aug 27;394(6696):909-13
pubmed: 9732876
Cell. 1998 Oct 30;95(3):379-91
pubmed: 9814708
Hum Reprod Update. 2015 Sep-Oct;21(5):616-26
pubmed: 26089403
BMC Bioinformatics. 2011 Aug 04;12:323
pubmed: 21816040
Science. 2018 Oct 19;362(6412):356-360
pubmed: 30237246
Bioinformatics. 2010 Jan 1;26(1):139-40
pubmed: 19910308
Nat Commun. 2017 Apr 21;8:15055
pubmed: 28429706
Cell Stem Cell. 2016 Jun 2;18(6):721-735
pubmed: 27257761
Mol Hum Reprod. 2018 Nov 1;24(11):543-555
pubmed: 30239859
Stem Cell Res Ther. 2013 Apr 24;4(2):36
pubmed: 23618383
Cell. 2003 May 30;113(5):631-42
pubmed: 12787504
Cell Stem Cell. 2017 Jun 1;20(6):858-873.e4
pubmed: 28457750
Nat Biotechnol. 2012 Feb 26;30(3):278-82
pubmed: 22371082
Mol Hum Reprod. 2018 May 1;24(5):233-243
pubmed: 29528446
Biol Reprod. 2017 Jan 1;97(6):850-861
pubmed: 29091993
Nat Commun. 2018 May 14;9(1):1873
pubmed: 29760424
Bioinformatics. 2013 Jan 1;29(1):15-21
pubmed: 23104886
Mech Dev. 2018 Feb;149:9-19
pubmed: 29129619
Stem Cells. 2021 May;39(5):551-563
pubmed: 33470497
Mol Hum Reprod. 2019 Sep 1;25(9):519-526
pubmed: 31211841
Stem Cells. 2015 Sep;33(9):2686-98
pubmed: 26108678
Nat Biotechnol. 2008 Mar;26(3):313-5
pubmed: 18278034
Stem Cells Dev. 2013 Dec 1;22(23):3141-55
pubmed: 23829223
Hum Reprod. 2011 Feb;26(2):316-22
pubmed: 21138906
Biol Reprod. 2017 Jun 1;96(6):1154-1166
pubmed: 28453617
Development. 2015 Sep 15;142(18):3151-65
pubmed: 26293300
Mol Hum Reprod. 2015 May;21(5):410-23
pubmed: 25634576
Development. 2009 Apr;136(8):1339-49
pubmed: 19279133
Cell Stem Cell. 2015 Aug 6;17(2):178-94
pubmed: 26189426
Nat Rev Mol Cell Biol. 2013 Jun;14(6):357-68
pubmed: 23673969
J Cell Sci. 2008 Mar 15;121(Pt 6):737-46
pubmed: 18322270