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Immortalized murine tenocyte cells: a novel and innovative tool for tendon research

Authors :
Gil Lola Oreff
Barbara Maurer
Ahmed N. ELKhamary
Iris Gerner
Veronika Sexl
Florien Jenner
Source :
Scientific Reports, Vol 13, Iss 1, Pp 1-10 (2023)
Publication Year :
2023
Publisher :
Nature Portfolio, 2023.

Abstract

Abstract Primary tenocytes rapidly undergo senescence and a phenotypic drift upon in vitro monolayer culture, which limits tendon research. The Ink4a/Arf locus encodes the proteins p16Ink4a/Arf and p14ARF (p19ARF in mice) that regulate cell cycle progression and senescence. We here established an immortalized cell line using tenocytes isolated from Ink4a/Arf deficient mice (Ink4a/Arf −/− ). These cells were investigated at three distinct time points, at low (2–5), intermediate (14–17) and high (35–44) passages. Wild-type cells at low passage (2–5) served as controls. Ink4a/Arf −/− tenocytes at all stages were comparable to wild-type cells regarding morphology, expression of tenogeneic genes (collagen type 1, 3 and 5, Scleraxis, Tenomodulin and Tenascin-C), and surface markers (CD29, CD44 and CD105) and form 3D tendon-like structures. Importantly, Ink4a/Arf −/− tenocytes maintained their phenotypic features and proliferation potential in culture for more than 40 passages and also following freeze–thaw cycles. In contrast, wild-type tenocytes underwent senescence starting in passage 6. These data define Ink4a/Arf −/− tenocytes as novel tool for in vitro tendon research and as valuable in vitro alternative to animal experiments.

Subjects

Subjects :
Medicine
Science

Details

Language :
English
ISSN :
20452322
Volume :
13
Issue :
1
Database :
Directory of Open Access Journals
Journal :
Scientific Reports
Publication Type :
Academic Journal
Accession number :
edsdoj.29c5e03e38841e08949311caea3ec42
Document Type :
article
Full Text :
https://doi.org/10.1038/s41598-023-28318-4