title: Immortalized Mouse Achilles Tenocytes Demonstrate Long-Term Proliferative Capacity While Retaining Tenogenic Properties.
abstract: Investigating the cellular processes underlying tendon healing can allow researchers to improve long-term outcomes after injury. However, conducting meaningful studies to uncover the injury healing mechanism at cellular and molecular levels remains challenging. This is due to the inherent difficulty in isolating, culturing, and expanding sufficient primary tenocytes, due to their limited proliferative capacity and short lifespan. In this study, we sought to establish a novel line of immortalized mouse Achilles tenocytes (iMATs) with primary tenocyte properties, but increased proliferative capacity suitable for extensive in vitro experimentation. We show that isolated primary mouse Achilles tenocytes (pMATs) can be effectively immortalized using a piggyBac transposon expressing SV40 large T antigen flanked by FLP recombination target site (FRT). The resulting iMATs exhibit markedly greater proliferation and survival, which can be reversed with FLP recombinase. Furthermore, iMATs express the same set of tendon-specific markers as that of primary cells, although in lower levels, and respond similarly to exogenous stimulation with bone morphogenetic protein 13 (BMP13) as has been previously reported with pMATs. Taken together, our results suggest that iMATs acquire long-term proliferative capacity while maintaining tenogenic properties. We believe that iMATs are a suitable model for studying not only the native cellular processes involved in injury and healing, but also potential therapeutic agents that may augment the stability of tendon repair.
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.
authors.forename: Sahitya K
authors.initials: SK
authors.lastname: Denduluri
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.
authors.forename: Bryan
authors.initials: B
authors.lastname: Scott
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Lamplot
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.|2 Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing, China .
authors.forename: Liangjun
authors.initials: L
authors.lastname: Yin
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.|2 Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing, China .
authors.forename: Zhengjian
authors.initials: Z
authors.lastname: Yan
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.|2 Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing, China .
authors.forename: Zhongliang
authors.initials: Z
authors.lastname: Wang
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.
authors.forename: Jixing
authors.initials: J
authors.lastname: Ye
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.|2 Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing, China .
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.|2 Ministry of Education Key Laboratory of Diagnostic Medicine, The Affiliated Hospitals of Chongqing Medical University , Chongqing, China .
authors.forename: Qiang
authors.initials: Q
authors.lastname: Wei
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.
authors.forename: Maryam K
authors.initials: MK
authors.lastname: Mohammed
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.
authors.forename: Rex C
authors.initials: RC
authors.lastname: Haydon
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.
authors.forename: Richard W
authors.initials: RW
authors.lastname: Kang
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.
authors.forename: Tong-Chuan
authors.initials: TC
authors.lastname: He
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.
authors.forename: Aravind
authors.initials: A
authors.lastname: Athiviraham
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.
authors.forename: Sherwin H
authors.initials: SH
authors.lastname: Ho
authors.affiliation: 1 Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Pritzker School of Medicine , Chicago, Illinois.
authors.forename: Lewis L
authors.initials: LL
authors.lastname: Shi
mesh_terms: D000125:Achilles Tendon / Q000166:cytology*; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D019485:Bone Morphogenetic Proteins / Q000494:pharmacology; D002461:Cell Line, Transformed; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004254:DNA Nucleotidyltransferases / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D060888:Real-Time Polymerase Chain Reaction; D000070916:Tenocytes / Q000166:cytology* / Q000187:drug effects
chemical_list: D000952:Antigens, Polyomavirus Transforming; D015415:Biomarkers; D019485:Bone Morphogenetic Proteins; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase