Do cord blood CD34 cells stay the same or change after expansion: A comprehensive analysis of gene expression results before and after seven days of cell culture

Abstract

IntroductionHematopoietic stem cell transplantation is used to treat hematopoietic malignancies and bone marrow failure syndromes. Due to the difficulties of using these cells isolated from the bone marrow, an additional source for receiving essential hematopoietic cells is umbilical cord blood. But the main problem with using the umbilical cord is its insufficient blood volume. The ex-vivo reproductive system of hematopoietic stem cells can overcome this problem that has the ability for Transplantation and hematopoiesis in the long term. This study aimed to evaluate the expression profile of CD133 + umbilical cord blood microRNAs in different stages of hematopoietic stem cells before and after ex-vivo proliferation. Materials and methodsThe expression profile of 1034 types of microRNA of CD34 + hematopoietic stem cells of the umbilical cord was analyzed before and after ex vivo proliferation. After isolating CD34 + cells from the umbilical cord blood of normal-born babies using MACS (Magnetic-activated cell sorting) column, these cells were cultured in a Stem span culture medium containing SCF, FLT3, and TPO cytokines in 24-well plates. The expression profile of microRNAs was investigated on days 0 and 7 days after cultivation by the Real-Time PCR method. ResultsThe results showed that the production of two-thirds of the micro-RNAs was reduced during the proliferation process. The micro-RNA expression of hematopoietic stem cell proliferation was also lower. At the same time, micro-RNA expression related to differentiation was higher. ConclusionThe observed reduction in miRNA expression may be attributed to enhanced differentiation through proliferation. Therefore, miRNAs appear to be a viable option for regulating the proliferation processes of Hematopoietic stem cell transplantation.