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Establishment and Characterization of SV40 T-Antigen Immortalized Porcine Muscle Satellite Cell.
Ni, Mengru; He, Jingqing; Li, Tao; Zhao, Gan; Ji, Zhengyu; Ren, Fada; Leng, Jianxin; Wu, Mengyan; Huang, Ruihua; Li, Pinghua; Hou, Liming.
Afiliación
  • Ni M; College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China.
  • He J; Institute of Swine Science, Nanjing Agricultural University, Nanjing 210095, China.
  • Li T; Key Laboratory of Pig Genetic Resources Evaluation and Utilization (Nanjing) of Ministry of Agriculture and Rural Affairs, Nanjing Agricultural University, Nanjing 210095, China.
  • Zhao G; College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China.
  • Ji Z; Institute of Swine Science, Nanjing Agricultural University, Nanjing 210095, China.
  • Ren F; Key Laboratory of Pig Genetic Resources Evaluation and Utilization (Nanjing) of Ministry of Agriculture and Rural Affairs, Nanjing Agricultural University, Nanjing 210095, China.
  • Leng J; College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China.
  • Wu M; Institute of Swine Science, Nanjing Agricultural University, Nanjing 210095, China.
  • Huang R; Key Laboratory of Pig Genetic Resources Evaluation and Utilization (Nanjing) of Ministry of Agriculture and Rural Affairs, Nanjing Agricultural University, Nanjing 210095, China.
  • Li P; College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China.
  • Hou L; Institute of Swine Science, Nanjing Agricultural University, Nanjing 210095, China.
Cells ; 13(8)2024 Apr 18.
Article en En | MEDLINE | ID: mdl-38667318
ABSTRACT
Muscle satellite cells (MuSCs) are crucial for muscle development and regeneration. The primary pig MuSCs (pMuSCs) is an ideal in vitro cell model for studying the pig's muscle development and differentiation. However, the long-term in vitro culture of pMuSCs results in the gradual loss of their stemness, thereby limiting their application. To address this conundrum and maintain the normal function of pMuSCs during in vitro passaging, we generated an immortalized pMuSCs (SV40 T-pMuSCs) by stably expressing SV40 T-antigen (SV40 T) using a lentiviral-based vector system. The SV40 T-pMuSCs can be stably sub-cultured for over 40 generations in vitro. An evaluation of SV40 T-pMuSCs was conducted through immunofluorescence staining, quantitative real-time PCR, EdU assay, and SA-ß-gal activity. Their proliferation capacity was similar to that of primary pMuSCs at passage 1, and while their differentiation potential was slightly decreased. SiRNA-mediated interference of SV40 T-antigen expression restored the differentiation capability of SV40 T-pMuSCs. Taken together, our results provide a valuable tool for studying pig skeletal muscle development and differentiation.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Diferenciación Celular / Antígenos Transformadores de Poliomavirus / Células Satélite del Músculo Esquelético Límite: Animals Idioma: En Revista: Cells Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: Suiza
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Diferenciación Celular / Antígenos Transformadores de Poliomavirus / Células Satélite del Músculo Esquelético Límite: Animals Idioma: En Revista: Cells Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: Suiza