Construction of pancreatic duodenal homeobox-1 recombinant retroviral vector and its stable expression in liver progenitor cells / 第二军医大学学报

ABSTRACT

Objective:To construct a recombinant retroviral vector expressing pancreatic duodenal homeobox-1(PDX-1) gene and express the PDX-1 expression cassette in liver progenitor cells(LEPCs),so as to study the conversion of hepatic cells to pancreatic-like cells.Methods: The full length of PDX-1 gene was amplified by PCR and subcloned into pMSCVpuro vector.The pMSCV PDX-1 puro was then introduced into Phoenix package cells and the cell culture supernatant was used for Ping-Pong infection of another packaging cell line PT67 to obtain stable virus-producing cell line.Results: The pMSCV PDX-1 puro vector was successfully constructed and transfected into PT67 cells.The viral supernatants of PT67 cells efficiently infected LEPCs in vitro and the infected LEPCs stably expressed PDX-1 gene.Conclusion: The pMSCV PDX-1 puro vector and PDX-1 expressing LEPCs have been successfully constructed,laying a basis for studying the transdifferentiation of liver progenitor cells to pancreas lineage cells.