World journal of gastroenterology · 2014
Reversible immortalization of human hepatocytes mediated by retroviral transfer and site-specific recombination.
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Meng Fan-Ying, Liu Li, Yang Feng-Hui, Li Chun-You, Liu Jun, Zhou Ping
Аннотация
AIM: To establish a method for the reversible immortalization of human hepatocytes, which may offer a good and safe source of hepatocytes for practical applications.
METHODS: We successfully isolated primary human hepatocytes from surgically resected liver tissue taken from a patient with liver hemangiomas. The freshly isolated cells were then immortalized with retroviral vector SSR#69 expressing simian virus 40 large T antigen (SV40T) and hygromycin-resistance genes flanked by paired loxP recombination targets.
RESULTS: The freshly isolated hepatocytes with high viability (85%) were successfully immortalized using retroviral gene transfer of SV40T. SV40T in the immortalized cells was then excised by Cre/loxP site-specific recombination. This cell population exhibited the characteristics of differentiated hepatocytes.
CONCLUSION: We successfully established reversibly immortalized human hepatocytes, which will provide an unlimited supply of cells for practical applications.
METHODS: We successfully isolated primary human hepatocytes from surgically resected liver tissue taken from a patient with liver hemangiomas. The freshly isolated cells were then immortalized with retroviral vector SSR#69 expressing simian virus 40 large T antigen (SV40T) and hygromycin-resistance genes flanked by paired loxP recombination targets.
RESULTS: The freshly isolated hepatocytes with high viability (85%) were successfully immortalized using retroviral gene transfer of SV40T. SV40T in the immortalized cells was then excised by Cre/loxP site-specific recombination. This cell population exhibited the characteristics of differentiated hepatocytes.
CONCLUSION: We successfully established reversibly immortalized human hepatocytes, which will provide an unlimited supply of cells for practical applications.
Предметные рубрики MeSH
Anti-Bacterial AgentsAntigens, Polyomavirus TransformingCell DifferentiationCell LineCell ProliferationCell SeparationCell SurvivalCell Transformation, ViralCinnamatesDrug Resistance, BacterialGenetic VectorsHepatocytesHumansHygromycin BIntegrasesRecombination, GeneticRetroviridaeSerum AlbuminSerum Albumin, HumanTime FactorsTransduction, Genetic