Investigating the effect of substitution of Acellular Wharton's Jelly for FBS on the megakaryocyte lineage specific markers of CD34+ cells
Abstract
Hematopoietic stem cells (HSCs CD34 +) have the ability to self-renewality. They are able to differentiate into all the different blood cells. Stem cell transplantation Hematopoiesis (HSCs) is a viable treatment for many cancers, malignant and non-malignant blood disorders, and immune system disorders. But the main disadvantage of using hematopoietic stem cells is the small number of these cells. Therefore, what is done to increase the number of HSCs is the proliferation of these cells in vitro. Animal serum-based culture medium, especially b fetal bovin serum (FBS), is used for cell proliferation. Since hematopoietic stem cells (HSCsCD34 +) are transplanted today, despite the removal of culture media and their materials from the cells, some of these materials are still injected into the transplant recipient. They accept that it is necessary to replace these environments with healthy ones, at least with human resources. One of the media that can be used as an alternative to animal serom is Warton Asellular Jelly (AWJ). Wharton's Acellular jelly contains a wide range of growth factors including: factor Platelet-derived growth (PDGF), epithelial growth factor (EGF), fibroblast growth factor (bFGF), acid fibroblast growth factor (aFGF), beta 1 metastatic growth factor β1-TGF), and insulin-like growth factor (I-IGF). There are receptors for these molecules on hematopoietic stem cells and precursors of blood receptors. All of these molecules affect cell proliferation and function and may alter proliferation relative to FBS.
Materials and Methods: UCB-derived CD34+ cell were Cultured in the presence of 20 ng/ml SCF and 100ng/ml TPO to evaluate the effect of replacing A Cellular Wharton's jelly with FBS on early differentiation into erythroid lineage in IMDM control groups containing 10% FBS and IMDM test group comprising 10%A cellular: Wharton's Jelly. The expression of (CD41) on the cell membrane surface was investigated by flow cytometry technique and two specific erythroid genes including GATA 2 and FLI1were analyzed by Real Time _PCR technique
Results: In this study, the expression of (CD41) in the presence of A Cellular Wharton's Jelly on the first and 3rd days was significantly increased compared to the control group. The results of PCR analysis showed that the expression of GATA2 & FLI1 genes on First and third days significantly increased