Isolation and Characterization of Highly Pure Type A Spermatogonia From Sterlet (
PLZF
fluorescence-activated cell sorting
germ stem cell
gonad
spermatogonia
sturgeon
Journal
Frontiers in cell and developmental biology
ISSN: 2296-634X
Titre abrégé: Front Cell Dev Biol
Pays: Switzerland
ID NLM: 101630250
Informations de publication
Date de publication:
2021
2021
Historique:
received:
08
09
2021
accepted:
08
11
2021
entrez:
27
12
2021
pubmed:
28
12
2021
medline:
28
12
2021
Statut:
epublish
Résumé
Sturgeons are among the most ancient linages of actinopterygians. At present, many sturgeon species are critically endangered. Surrogate production could be used as an affordable and a time-efficient method for endangered sturgeons. Our study established a method for identifying and isolating type A spermatogonia from different developmental stages of testes using flow cytometric cell sorting (FCM). Flow cytometric analysis of a whole testicular cell suspension showed several well-distinguished cell populations formed according to different values of light scatter parameters. FCM of these different cell populations was performed directly on glass slides for further immunocytochemistry to identify germ cells. Results showed that the cell population in gate P1 on a flow cytometry plot (with high forward scatter and high side scatter parameter values) contains the highest amount of type A spermatogonia. The sorted cell populations were characterized by expression profiles of 10 germ cell specific genes. The result confirmed that setting up for the P1 gate could precisely sort type A spermatogonia in all tested testicular developmental stages. The P2 gate, which was with lower forward scatter and side scatter values mostly, contained type B spermatogonia at a later maturing stage. Moreover, expressions of
Identifiants
pubmed: 34957105
doi: 10.3389/fcell.2021.772625
pii: 772625
pmc: PMC8708567
doi:
Types de publication
Journal Article
Langues
eng
Pagination
772625Informations de copyright
Copyright © 2021 Xie, Tichopád, Kislik, Langerová, Abaffy, Šindelka, Franěk, Fučíková, Steinbach, Shah, Šauman, Chen and Pšenička.
Déclaration de conflit d'intérêts
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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