| First Author | Oreff GL | Year | 2023 |
| Journal | Sci Rep | Volume | 13 |
| Issue | 1 | Pages | 1566 |
| PubMed ID | 36709227 | Mgi Jnum | J:334801 |
| Mgi Id | MGI:7432002 | Doi | 10.1038/s41598-023-28318-4 |
| Citation | Oreff GL, et al. (2023) Immortalized murine tenocyte cells: a novel and innovative tool for tendon research. Sci Rep 13(1):1566 |
| abstractText | 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 p16(Ink4a/Arf) and p14(ARF) (p19(ARF) 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. |
