(中国肉类食品综合研究中心 北京食品科学研究院 北京 100068)
(China Meat Research Center， Beijing Institute of Food Science， Beijing 100068)
Objective: Construction of immortalized seed cells is an efficient and sustainable way to solve the problem of seed cells of cultured meat. Methods: Based on the current industrialized production of cultured meat， the source of seed cells needs to be solved urgently， and it is also a technical bottleneck in the large-scale production of cell-cultured meat. In this study， based on porcine myoblasts， the inducible vector was constructed by telomerase reverse transcriptase， and the cell induction was verified by immunofluorescence labeling， quantitative PCR， and karyotype analysis after nucleofection into the cells. Results: showed that: the construction of the immortalized cell induction vector had high sequence integrity and no mutation； the vector was successfully transferred into the cells and expressed efficiently， and one induction positive cell line was obtained； no uncontrollable mutation occurred in the chromosomal karyotype after transfection； The expression of telomerase reverse transcriptase in the cells was significantly higher than that in the control group； more importantly， the number of passages of induced positive cells exceeded the 'Hayflick＇ proliferation limit， achieving more than 200 passages. Conclusion: we constructed the immortalization-inducing carrier of porcine myoblasts， described the stability of cells after induction， which will have a broad application prospect in the production process of cultured meat.