Immortalization of primary sheep embryo kidney cells.
Cell immortalization
Epithelial kidney cells
SV40 T antigen
Transfection
Journal
In vitro cellular & developmental biology. Animal
ISSN: 1543-706X
Titre abrégé: In Vitro Cell Dev Biol Anim
Pays: Germany
ID NLM: 9418515
Informations de publication
Date de publication:
Jan 2021
Jan 2021
Historique:
received:
06
03
2020
accepted:
13
10
2020
pubmed:
9
1
2021
medline:
25
9
2021
entrez:
8
1
2021
Statut:
ppublish
Résumé
Sheep primary epithelial cells are short-lived in cell culture systems. For long-term in vitro studies, primary cells need to be immortalized. This study aims to establish and characterize T immortalized sheep embryo kidney cells (TISEKC). In this study, we used fetal lamb kidneys to derive primary cultures of epithelial cells. We subsequently immortalized these cells using the large T SV40 antigen to generate crude TISEKC and isolate TISEKC clones. Among numerous clones of immortalized cells, the selected TISEKC-5 maintained active division and cell growth over 20 passages but lacked expression of the oncogenic large T SV40 antigen. Morphologically, TISEKC-5 maintained their epithelial aspect similar to the parental primary epithelial cells. However, their growth properties showed quite different patterns. Crude TISEKC, as well as the clones of TISEKC proliferated highly in culture compared to the parental primary cells. In the early passages, immortalized cells showed heterogeneous polyploidy but in the late passages the karyotype of immortalized cells became progressively stable, identical to that of the primary cells, because the TISEKC-5 cell line has lost the large SV40 T antigen expression, this cell line is a valuable tool for veterinary sciences and biotechnological productions.
Identifiants
pubmed: 33415664
doi: 10.1007/s11626-020-00520-y
pii: 10.1007/s11626-020-00520-y
doi:
Substances chimiques
Antigens, Viral, Tumor
0
DNA, Viral
0
Vimentin
0
Serum Albumin, Bovine
27432CM55Q
Keratins
68238-35-7
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
76-85Subventions
Organisme : Communauté Université Grenoble Alpes
ID : FR-2015
Organisme : Institut National de la recherche Agronomique
ID : SA 2010-2015
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