Cultivation and differentiation of human umbilical cord-derived mesenchymal stem cells treated with amphotericin B / 中国组织工程研究

ABSTRACT

BACKGROUND:High-quality and efficient umbilical cord-derived mesenchymal stem cells could be obtained by establishing and improving the method to avoid fungal contamination and by effectively decreasing pol ution rate of isolated culture during umbilical cord col ection. OBJECTIVE:To explore the effects of amphotericin B on growth and differentiation potential of umbilical cord-derived mesenchymal stem cells. METHODS:Umbilical cord-derived mesenchymal stem cells were separated from healthy ful-termed delivery fetus using col agenase digestion method and treated with amphotericin B. Subsequently, umbilical cord-derived mesenchymal stem cells were amplified and cultured in vitro with MesenPRO RSTM medium. The third passage of umbilical cord-derived mesenchymal stem cells in logarithmic phase was obtained to analyze their morphology, proliferation and immunophenotype, and induced to differentiate into osteoblasts and adipocytes in vitro. RESULTS AND CONCLUSION:After treatment with amphotericin B, umbilical cord-derived mesenchymal stem cells were successful y isolated and cultured in vitro. Flow cytometry results revealed that human umbilical cord-derived mesenchymal stem cells strongly expressed CD44, CD105 and CD73, CD90, and negatively expressed HLA-DR, CD29, CD31, and CD34. Amphotericin B-treated human umbilical cord-derived mesenchymal stem cells can stil differentiate into adipocytes and osteoblasts in vitro.