Construction of MafA recombinant retroviral vector and its stable expression in liver epithelial progenitor cells / 第二军医大学学报

ABSTRACT

Objective: To construct a recombinant retroviral vector harboring MafA and to establish a liver epithelial progenitor cell (LEPC) line for stable expression of MafA. Methods: MafA was amplified by PCR and suncloned into pBMN-Z-IRES-Neo vector to obtain pBMH-MafA-Neo vector. After introducing the pBMN-MafA-Neo into Phoenix package cells, the cell culture supernatant was used to infect LEPCs. LEPCs stably expressing MafA gene were screened out. RT-PCR was used to detect the influence of MafA on the phenotype of LEPCs. Results: We successfully constructed pBMH-MafA-Neo vector and obtained LEPCs which stably expressed MafA. Expression of GK and GLUT2 genes in LEPCs-MafA cells was higher than that in the LEPCs. Conclusion: We have successfully obtained LEPCs which can stably express MafA,laying a basis for studying the differentiation of LEPCs into pancreas cells.