Isolation and purification of human umbilical cord blood mesenchymal stem cells in vitro / 中华内分泌外科杂志

ABSTRACT

Objective To induce the differentiation of human umbilical cord blood MSCs into osteoblasts in vitro,and to study the method of inducing MSCs to differentiate into osteoblasts under specific microenvironment.Methods MSCs was obtained from human umbilical vein,and isolated by density gradient method.The morphological changes of MSCs were observed by using dexamethasone,beta sodium phosphate and vitamin C.The proliferation and differentiation of MSC in cord blood were studied by means of optical microscope,transmission electron microscope and alkaline phosphatase staining.The expression of bone morphogenetic protein 2 mRNA in human umbilical cord blood MSCs after osteogenic was inducted by RT-PCR.Results After the umbilical cord blood MSCs were differentiated into osteoblastic cells in microenvironment,fusiform cells became polygonal,irregular shape,local cells presented overlapping growth.After 10 days,the cells gradually presented square,crystal particles of high refraction,and began to show the characteristics of osteoblasts.The expression of bone morphogenetic protein 2 mRNA was positive in alkaline phosphatase staining and alizarin red staining.Conclusion Human umbilical cord blood MSCs can be induced into osteoblasts in vitro,which is an ideal seed cell for bone tissue engineering.