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.
Highlights
The chicken is an important agricultural animal throughout the world and is a significant non-mammalian vertebrate model for developmental biology, virology and immunology
Heat treatment (HT) caused a loss of telomerase activity in the immortalized chicken preadipocyte 1 (ICP1) and immortalized chicken preadipocyte 2 (ICP2) samples (Fig 3B). These results indicated that these two cell lines gained and maintained telomerase activity and that chicken telomerase reverse transcriptase (chTERT), not chicken telomerase RNA (chTR), was the rate-limiting factor for chicken preadipocyte immortalization
We successfully established two immortalized chicken preadipocyte lines (ICP1 and ICP2) through the introduction of chicken TERT and TR genes. These two cell lines underwent more than 100 cumulative population doublings (PDs) (Fig 1), which far exceeded the maximum in vitro life span of the primary chicken preadipocytes in our studies
Summary
The chicken is an important agricultural animal throughout the world and is a significant non-mammalian vertebrate model for developmental biology, virology and immunology. Chickens show high glycemia and low sensitivity to exogenous insulin ( in adipose tissues), making them a potential model for studies on human obesity, insulin resistance and type 2 diabetes [1,2,3,4,5]. In the broiler chicken industry, excessive fat deposition has been a growing concern that urgently needs to be addressed, because it reduces carcass yield and feed efficiency and causes processing difficulties and environmental pollution. Immortalization of chicken preadipocytes analysis, decision to publish, or preparation of the manuscript
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