Isolation and characterization of mesenchymal stem cells derived from whole human umbilical cord by applying a direct explant technique

Objective To establish a technique for isolating mesenchymal stem cells from umbilical cord and their biological characteristics.Methods Mesenchymal stem cells(MSCs) were isolated from umbilical cord(UC) by applying a explant technique and explanding cultured in vitro.Growth curves were drawn,and The phenotypes and cell cycle were evaluated by flow cytometry.UC-MSCs were induced to differentiate into adipocytes,osteocytes and chondrocytes in special differentiation condition.Mutidifferentiation related genes and stem cell-related transcription factors,Nanog,Oct-4 Sox-2 were detected by TR-PCR.Results UC-MSCs were isolated using a explant technique,and characteristic of plastic adherence and fibroblast-like morphology.the adherent cells displayed an abundant presence of CD73,CD90,CD105 and absence of CD34,CD45,HLA-DR.Cell cycle showed that there were percentages of stem cells as G0/G1 81.56 % and S+G2 + M 18.44% respectively.When cultured in differentiation media,they differentiated into adipocytes,osteocytes and chondrocytes.RT-PCR reactions confirmed that their mutidifferentiation related genes were positive.Moreover,stem cell-related transcription factors,Nanog,Oct-4 Sox-2 were positively expressed in UC-MSCs.Conclusions The explant technique is a feasible method to obtain MSCs from whole human umbilical cord,and UC can be considered as a novel and convenient source of adult MSCs displaying primitive pluripotent sten cells and high expansion potential.