Generation of a human haploid neural stem cell line for genome-wide genetic screening.
Cell therapy
Extended pluripotency
Genetic screening
Haploid
Neural stem cells
Journal
World journal of stem cells
ISSN: 1948-0210
Titre abrégé: World J Stem Cells
Pays: United States
ID NLM: 101535826
Informations de publication
Date de publication:
26 Jul 2023
26 Jul 2023
Historique:
received:
21
03
2023
revised:
01
06
2023
accepted:
21
06
2023
medline:
7
8
2023
pubmed:
7
8
2023
entrez:
7
8
2023
Statut:
ppublish
Résumé
Haploid embryonic stem cells (haESCs) have been established in many species. Differentiated haploid cell line types in mammals are lacking due to spontaneous diploidization during differentiation that compromises lineage-specific screens. To derive human haploid neural stem cells (haNSCs) to carry out lineage-specific screens. Human haNSCs were differentiated from human extended haESCs with the help of Y27632 (ROCK signaling pathway inhibitor) and a series of cytokines to reduce diploidization. Neuronal differentiation of haNSCs was performed to examine their neural differentiation potency. Global gene expression analysis was con-ducted to compare haNSCs with diploid NSCs and haESCs. Fluorescence activated cell sorting was performed to assess the diploidization rate of extended haESCs and haNSCs. Genetic manipulation and screening were utilized to evaluate the significance of human haNSCs as genetic screening tools. Human haESCs in extended pluripotent culture medium showed more compact and smaller colonies, a higher efficiency in neural differentiation, a higher cell survival ratio and higher stability in haploidy maintenance. These characteristics effectively facilitated the derivation of human haNSCs. These human haNSCs can be generated by differentiation and maintain haploidy and multipotency to neurons and glia in the long term This is the first report of a generated human haploid somatic cell line with a complete genome, proliferative ability and neural differentiation potential that provides cell resources for recessive inheritance and drug targeted screening.
Sections du résumé
BACKGROUND
BACKGROUND
Haploid embryonic stem cells (haESCs) have been established in many species. Differentiated haploid cell line types in mammals are lacking due to spontaneous diploidization during differentiation that compromises lineage-specific screens.
AIM
OBJECTIVE
To derive human haploid neural stem cells (haNSCs) to carry out lineage-specific screens.
METHODS
METHODS
Human haNSCs were differentiated from human extended haESCs with the help of Y27632 (ROCK signaling pathway inhibitor) and a series of cytokines to reduce diploidization. Neuronal differentiation of haNSCs was performed to examine their neural differentiation potency. Global gene expression analysis was con-ducted to compare haNSCs with diploid NSCs and haESCs. Fluorescence activated cell sorting was performed to assess the diploidization rate of extended haESCs and haNSCs. Genetic manipulation and screening were utilized to evaluate the significance of human haNSCs as genetic screening tools.
RESULTS
RESULTS
Human haESCs in extended pluripotent culture medium showed more compact and smaller colonies, a higher efficiency in neural differentiation, a higher cell survival ratio and higher stability in haploidy maintenance. These characteristics effectively facilitated the derivation of human haNSCs. These human haNSCs can be generated by differentiation and maintain haploidy and multipotency to neurons and glia in the long term
CONCLUSION
CONCLUSIONS
This is the first report of a generated human haploid somatic cell line with a complete genome, proliferative ability and neural differentiation potential that provides cell resources for recessive inheritance and drug targeted screening.
Identifiants
pubmed: 37545755
doi: 10.4252/wjsc.v15.i7.734
pmc: PMC10401418
doi:
Types de publication
Journal Article
Langues
eng
Pagination
734-750Informations de copyright
©The Author(s) 2023. Published by Baishideng Publishing Group Inc. All rights reserved.
Déclaration de conflit d'intérêts
Conflict-of-interest statement: The authors declare having no conflicts of interest.
Références
Stem Cell Reports. 2020 Jul 14;15(1):185-197
pubmed: 32502463
J Comp Neurol. 2000 Sep 11;425(1):58-69
pubmed: 10940942
Science. 2009 Oct 16;326(5951):430-3
pubmed: 19833967
Cell Stem Cell. 2014 Oct 2;15(4):488-497
pubmed: 25280221
DNA Res. 2014 Oct;21(5):541-54
pubmed: 24939742
Cell. 2014 Sep 11;158(6):1254-1269
pubmed: 25215486
Annu Rev Cell Dev Biol. 2014;30:705-22
pubmed: 25288120
Nature. 2012 Oct 18;490(7420):407-11
pubmed: 23023130
Cell Res. 2020 Sep;30(9):810-813
pubmed: 32152419
Nat Commun. 2021 Nov 18;12(1):6718
pubmed: 34795250
Cell Death Dis. 2022 Mar 30;13(3):284
pubmed: 35354792
Cell Res. 2020 Apr;30(4):356-359
pubmed: 31853003
Toxicol Res (Camb). 2015 Mar 1;4(2):191-202
pubmed: 25893090
Development. 2018 Jun 11;145(11):
pubmed: 29784672
Nature. 2009 Apr 9;458(7239):766-70
pubmed: 19252478
Nature. 2013 Dec 12;504(7479):282-6
pubmed: 24172903
Cell Prolif. 2022 Apr;55(4):e13209
pubmed: 35274380
Cell. 2012 Apr 27;149(3):605-17
pubmed: 22541431
Cell Res. 2013 Oct;23(10):1187-200
pubmed: 23856644
Cell Prolif. 2020 Feb;53(2):e12754
pubmed: 31916359
J Stroke Cerebrovasc Dis. 2013 May;22(4):304-8
pubmed: 22078778
Cell Prolif. 2022 Nov;55(11):e13317
pubmed: 35880490
Cell Res. 2016 Jun;26(6):743-6
pubmed: 27185278
Cell. 2016 Jan 14;164(1-2):279-292
pubmed: 26771496
Dev Cell. 2019 Dec 16;51(6):698-712.e8
pubmed: 31846649
Genome Biol. 2006;7 Suppl 1:S4.1-9
pubmed: 16925838
Cell Death Dis. 2020 Jan 23;11(1):52
pubmed: 31974372
Curr Drug Targets. 2012 May;13(5):602-6
pubmed: 22300026
Nature. 2011 Sep 07;479(7371):131-4
pubmed: 21900896
Cell Stem Cell. 2011 Jan 7;8(1):46-58
pubmed: 21211781
Protein Cell. 2019 Jan;10(1):20-30
pubmed: 29948855
Cell. 2005 Aug 12;122(3):473-83
pubmed: 16096065
Nat Biotechnol. 2001 Dec;19(12):1129-33
pubmed: 11731781
Science. 2022 Aug 26;377(6609):967-975
pubmed: 36007034
Cell. 2019 Oct 17;179(3):687-702.e18
pubmed: 31626770
Cell Stem Cell. 2011 Dec 2;9(6):563-74
pubmed: 22136931
Cell Rep. 2017 Aug 29;20(9):2227-2237
pubmed: 28854370
Nat Commun. 2021 May 21;12(1):3017
pubmed: 34021145
Nature. 2016 Apr 7;532(7597):107-11
pubmed: 26982723
Nat Protoc. 2009;4(5):789-98
pubmed: 19528954
Biol Direct. 2014 Oct 14;9(1):20
pubmed: 25319552
Cell. 2017 Apr 6;169(2):243-257.e25
pubmed: 28388409
Brain Pathol. 2006 Apr;16(2):132-42
pubmed: 16768754
Cell. 2021 Jun 24;184(13):3589
pubmed: 34171321
J Neurosci. 2012 Apr 11;32(15):5039-53
pubmed: 22496550
Protein Cell. 2020 Jan;11(1):23-33
pubmed: 31004328
Stem Cells Transl Med. 2021 May;10(5):743-755
pubmed: 33511777
Cell Stem Cell. 2014 Mar 6;14(3):404-14
pubmed: 24360884
Genes Dev. 2008 Jan 15;22(2):152-65
pubmed: 18198334