Purpose: To investigate the transfer efficiency and transient expression of plasmid vector coding enhanced green fluorescent protein (EGFP)gene pEGFP-N1 which was transferred into primary cultured human fetal retinal progenitor cells. To build up a tracking method for study of retinal progenitor cells transplantation.
Methods: pEGFP-N1 plasmid was amplified in E.coli, primary cultured human fetal retinal progenitor cells transferred with pEGFP-N1 by means of CaCl2. The transfer efficiency was evaluated by flow cytometry. Transient expression in vitro was observed by fluorescent microscope and laser scanning confocal microscope.
Results: The transfer efficiency of pEGFP-N1 into primary cultured human fetal retinal progenitor cells could maximumly reach to 32.45% during 48 hours. There was obvious expression of pEGFP-N1 in vitro at 48th hour after transferring.
Conclusion: pEGFP-N1 is an ideal transient expression vector for primary cultured human fetal retinal progenitor cells in vitro. It is a useful tracer for transplantation of retinal progenitor cells.