Propagation of porcine spermatogonial stem cells in serum-free culture conditions using knockout serum replacement.
Doom pig
immunofluorescence
qPCR
serum-free culture
spermatogonial stem cell
Journal
Reproduction in domestic animals = Zuchthygiene
ISSN: 1439-0531
Titre abrégé: Reprod Domest Anim
Pays: Germany
ID NLM: 9015668
Informations de publication
Date de publication:
Feb 2023
Feb 2023
Historique:
revised:
30
09
2022
received:
12
08
2022
accepted:
09
10
2022
pubmed:
13
10
2022
medline:
8
2
2023
entrez:
12
10
2022
Statut:
ppublish
Résumé
In vitro culture and expansion of spermatogonial stem cells (SSCs) is an essential prerequisite to enhancing livestock productivity through SSC transplantation. Most of the culture media have been observed to be supplemented with serum. However, the use of serum in culture media may exert detrimental effects on SSC maintenance in vitro. An attempt was made to culture SSCs by replacing serum with 5% 'Knockout Serum Replacement (KSR)' in Doom pig (Sus domesticus), one of the valued indigenous germplasm of North-East India. Testes from 7 to 15 days old piglets were used for isolation, enrichment and in vitro culture of putative SSCs using serum-based and serum-free culture media. The cells were characterized for SSC-specific pluripotent markers expression by immunofluorescence staining and quantitative real-time PCR. The diameter and number of SSC colonies were recorded on days 9, 20 and 30 of culture. Similar morphologies of the SSC colonies were observed in both serum-based and serum-free culture conditions. Colony diameter and colony number were non-significantly higher in serum-free than serum-based media. The cells from both the culture conditions showed high alkaline phosphatase activity. The expression of SSC-specific pluripotent markers was observed in immunofluorescence and quantitative real-time PCR study. The present study revealed that SSCs from porcine species could be maintained in vitro for up to 30 days in serum-free culture using 5% KSR, which is believed to be a promising protein source for improving livestock production, and health care along with their conservation.
Substances chimiques
Culture Media
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
219-229Subventions
Organisme : Department of Biotechnology Govt of India
Commentaires et corrections
Type : ErratumIn
Informations de copyright
© 2022 Wiley-VCH GmbH.
Références
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