مقالات پذیرفته شده در سومین کنگره بین المللی زیست پزشکی
Effect of platelet lysate replacement with fetal bovine serum on the expansion and early stages of erythroid differentiation of Hematopoietic stem cells for Hematopoietic tissue cell therapy
Effect of platelet lysate replacement with fetal bovine serum on the expansion and early stages of erythroid differentiation of Hematopoietic stem cells for Hematopoietic tissue cell therapy
Majid Zamani,1Yoda Yaghoubi,2Mehdi Yousefi,3,*AliAkbar Movasaghpour,4Amir Mehdizadeh,5Alireza Pishgahi,6
1. Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran 2. Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran 3. Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran 4. Hematology and Oncology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran 5. Endocrine Research Center, Tabriz university of Medical Sciences, Tabriz, Iran 6. Physical Medicine and Rehabilitation research Center, Tabriz University of Medical Science, Iran
Introduction: Impaired hematopoietic tissue function may be occurred due to a variety of reasons such as tumors or leukemia. Stem cell therapy is one of the approaches for the treatment of these disorders to produce hematopoietic tissue or mature cells to be replaced with blood cells. Recently, hematopoietic tissue and red blood cells production have been used in clinical studies as a novel therapeutic approach. The production of these cells mainly relies on the fetal bovine serum (FBS) containing or serum-free media which may arise the xenoviruses transmission concerns and high cost. One suitable alternative for FBS is platelet lysate, which contains growth factors and cytokine and can promote cell growth, proliferation and differentiation. The aim of this study was to assess the effect of platelet lysate on early differentiation of erythrocytes.
Methods: Heparinized umbilical cord blood (CB) was obtained at the end of full-term deliveries. Mononuclear cells (MNCs) were isolated from CB using ficoll hypaque (density, 1077). The CD34+/MNC fraction was isolated with superparamagnetic microbeads selection using high-gradient magnetic field and mini MACS columns. Cells were then cultured (1×104 Cell/ml) for 3 days in IMDM medium containing 10% platelet lysate and SCF, Tpo, Epo, FL. FBS containing medium was also considered as control. After 3 days, GPA CD34, CD71, and CD36 expression were evaluated using MAX technique. The number of nucleated cells was also considered as proliferation marker.
Results: Immunological characterization showed expression of CD36 (5% and 3%), CD71 (55% and 51%), CD34 (56% and 61%) at the day 3 in platelet lysate and FBS containing media, respectively. GPA expression in both groups was <1% at day 3. Additionally, in platelet lysate and FBS containing media a mean of 8 and 7 fold cell expansion were observed, respectively.
Conclusion: Higher expression of CD71 (transferrin receptor) and lower expression of CD34 in platelet lysate containing media indicated more differentiation of cells in early erythroid stages. In addition Lack of GPA expression indicates the early stages of differentiation of these cells. As well as the higher proliferation rate in platelet lysate containing media showed the more effect of platelet lysate on cell proliferation. The results of this study indicate that platelet lysate is a suitable alternative for FBS to produce red blood cells and hematopoietic tissue without xenoviruses transmission and immune reactions.