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