Immortalization of chicken preadipocytes by retroviral transduction of chicken TERT and TR

PLoS One. 2017 May 9;12(5):e0177348. doi: 10.1371/journal.pone.0177348. eCollection 2017.

Abstract

The chicken is an important agricultural animal and model for developmental biology, immunology and virology. Excess fat accumulation continues to be a serious problem for the chicken industry. However, chicken adipogenesis and obesity have not been well investigated, because no chicken preadipocyte cell lines have been generated thus far. Here, we successfully generated two immortalized chicken preadipocyte cell lines through transduction of either chicken telomerase reverse transcriptase (chTERT) alone or in combination with chicken telomerase RNA (chTR). Both of these cell lines have survived >100 population doublings in vitro, display high telomerase activity and have no sign of replicative senescence. Similar to primary chicken preadipocytes, these two cell lines display a fibroblast-like morphology, retain the capacity to differentiate into adipocytes, and do not display any signs of malignant transformation. Isoenzyme analysis and PCR-based analysis confirmed that these two cell lines are of chicken origin and are free from inter-species contamination. To our knowledge, this is the first report demonstrating the generation of immortal chicken cells by introduction of chTERT and chTR. Our established chicken preadipocyte cell lines show great promise as an in vitro model for the investigation of chicken adipogenesis, lipid metabolism, and obesity and its related diseases, and our results also provide clues for immortalizing other avian cell types.

MeSH terms

  • Adipocytes / metabolism
  • Animals
  • Cell Line, Transformed
  • Chickens
  • Cloning, Molecular
  • Genetic Vectors
  • Polymerase Chain Reaction
  • RNA / genetics
  • Retroviridae / genetics*
  • Telomerase / genetics*
  • Transduction, Genetic*

Substances

  • RNA
  • Telomerase

Grants and funding

This work was supported by the National High-tech R&D Program (863 Program) (No. 2013AA102501), the National Natural Science Foundation of China (No. 30972086), the National Basic Research Program of China (973 Program) (No. 2009CB941604) and the China Agricultural Research System (No. CARS-42). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.