Preparation of pigment epithelial-derived factor-modified human umbilical cord-derived mesenchymal stem cells

Abstract

AIM: To construct the recombined pLenti-CMV-PEDF-EYFP vector to be infected into human umbilical cord-derived mesenchymal stem cells(hUCMSCs), and then to identify the PEDF expression in hUCMSCs and the cell viability of hUCMSCs after infection with the recombined lentivirus. METHODS: The recombinant pLenti-CMV-PEDF-EYFP vector was constructed with pLenti-CMV-hChR2-EYFP and PEDF gene, and then infected into hUCMSCs to obtain the PEDF-hUCMSCs. The expression of PEDF in hUCMSCs was identified by confocal microscopy and ELISA. The Cell Counting Kit(CCK8)was used to assess the cell viability of PEDF-hUCMSCs. The influences of pLenti-CMV-PEDF-EYFP infection on the passage and phenotypes of hUCMSCs were assessed by microscope and flow cytometry. RESULTS: The recombinant pLenti-CMV-PEDF-EYFP was successfully constructed and efficiently infected into hUCMSCs. The expression of PEDF was positively detected in the PEDF-hUCMSCs. No significant difference was observed on cell viability between PEDF-modified hUCMSCs and hUCMSCs(P>0.05). The infection of pLenti-CMV-PEDF-EYFP had no obvious influences on the proliferation, morphology and phenotypes of hUCMSCs. CONCLUSION: PEDF was expressed effectively in the hUCMSCs modified with the recombinant lentivirus. The preliminary results from this study provide more evidences for further study of using PEDF-hUCMSCs to treat retinitis pigmentosa.

Keywords

• pigment epithelial-derived factor
• lentiviral vector
• human umbilical cord-derived mesenchymal stem cell
• human umbilical cord-derived mesenchymal stem cell