Creation of a Stable P19 Cell Line Producing PTS2-EGFP

Abstract

Background: P19 cells are mouse embryonic carcinoma cells which contain pluripotent ability, like stem cells, to differentiate into different cell lines. There are several properties for this cell line that make it a valuable cell model for study of developmental stages. Methods: At the first step, PTS2-EGFP coding sequence which was cloned in pUcD2.hygro vector was used for transfection in to P19 cells. As the plasmid contained hygromycin (Hygror) resistance gene, stable cells were selected using hygromycin as an antibiotic. Stable transformed cells were characterized by RT-PCR and immunostaining analyses. Findings: RT-PCR results indicated EGFP was expressed in these cells. Moreover immunocytochemical analysis of the cells confirmed the preserved pluripotency states of transfected cells. Conclusion: As P19 Cells are able to differentiate into several cell lines, using this stable cell line we will able to chase the molecular kinetics of peroxisomes. Key words: Lipofection, P19, Plasmid, RT-PCR, peroxisome.