An efficient method for isolation, characterization and immunophenotypic analysis of human umbilical cord blood derived mesenchymal stem cells in vitro

Abstract EN

Background and Objective Umbilical cord blood (UCB) is an interesting source of mesenchymal stem cells (MSCs) for gene therapy, cell transplantation and cell therapy.

This study is aimed to isolate, characterize and immunophenotypic analysis of human UCB- derived MSCs by using a simplified technique.

Methods Cord blood was collected after normal delivery of placenta by puncturing umbilical cord veins.

The mononucleated cells (MNCs) was isolated after gradient centrifugation and cultured in Iscove,s Modified Dulbecco Medium (IMDM) supplemented with 10% Fetal calf serum (FCS).

Cultures were maintained at 37C°,5% CO2 for two weeks.

Then immunophenotypic analysis which was performed with CD90, CD71, CD34 and HLA-DR.

Results: The MSCs derived from MNCs appeared like the fibroblast cell and these cells were extensively expanded in culture.

Immunocytochemistry staining indicated that UCB-derived MSCs were positive for CD90, CD71 and negative for CD34, HLA-DR.

Conclusion: The result of the present study indicate that using this method can result in isolation of homogenous population of UCB- MSCs.