Guoji Yanke Zazhi (Jul 2017)

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

  • Hui Wen,
  • Yun Zhang,
  • Xi-Luan Ji,
  • Xiao-Lei Liu,
  • Shun Yang,
  • Zhao-Xia Luo,
  • Liang Xie,
  • Wen-Wen Zou,
  • Jie-Ming Li,
  • Shu Jiang

DOI
https://doi.org/10.3980/j.issn.1672-5123.2017.7.07
Journal volume & issue
Vol. 17, no. 7
pp. 1226 – 1231

Abstract

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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.

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