Investigating the effect of substitution of Platelet Lysate for FBS on the Erythroid related differentiation and proliferation pattern of CD34+ cells following co-culture with Mesenchymal Stem Cells

Abstract

Red blood cell transfusion is a supportive and common treatment in surgical care, trauma, treatment of malignancies and leukemia, and is especially used in bleeding anemia. Due to the limitation of blood resources, the possibility of disease transmission, immune reactions and the presence of rare blood groups, the production of these cells seems to be an important necessity. Production of red blood cells in the in vitro as an alternative to blood transfusions has attracted the attention of many scientists. This blood source can be a suitable alternative to blood transfusions, especially for rare blood groups, as well as a suitable source without the worry of transmitting bloodstream viruses such as HIV. cell cultures require culture media that are supplemented with FBS that can transmit animal xenoviruses in addition to harming animals and ethical problems or use serum-free mediums that have a high cost for culturing cells in high volume. One of the suitable alternatives to enrich the culture medium is the use of alternatives such as platelet lysate which, in addition to having high growth factor and low cost of production, can provide a suitable substrate for cell culture. Material and Method: In this study, 3-step culture was used for proliferation and differentiation of umbilical cord blood CD34 + stem cells to produce erythrocytes. Cell proliferation and expansion were assessed using cell counts on different days. Cells were cultured in platelet lysate-enriched medium for 15 days and on day 8 after culturing, erythroid differentiation genes were analyzed by Realtime-PCR. The expression levels of CD71 and Glycophorin A markers were evaluated by flow cytometry on days 0, 7 and 15. Result: Maximum proliferation of hematopoietic cells was observed on day 15. Expression of GATA1, NFE2, Globin β, Globin γ genes was significantly increased and GATA2 gene expression was decreased but not significant. Surveys of surface markers showed greater expression of erythroid markers and greater differentiation of cells in platelet lysate-containing medium.