Zamani Majid, Yaghoubi Yoda, Naimi Adel, Hassanzadeh Ali, Pourakbari Ramin, Aghebati-Maleki Leili, Motavalli Roza, Aghlmandi Afsoon, Mehdizadeh Amir, Nazari Mehdi, Yousefi Mehdi, Movassaghpour Ali Akbar
Department of Medical Laboratory Sciences, Faculty of Allied Medicine, Gonabad University of Medical Sciences, Gonabad, Iran.
Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Adv Pharm Bull. 2021 Feb;11(2):335-342. doi: 10.34172/apb.2021.031. Epub 2020 Feb 29.
Transfusion of red blood cells (RBCs) is a supportive and common treatment in surgical care, trauma, and anemia. However, in vivo production of RBC seems to be a suitable alternative for blood transfusions due to the limitation of blood resources, the possibility of disease transmission, immune reactions, and the presence of rare blood groups. Cell cultures require serum-free or culture media supplemented with highly expensive animal serum, which can transmit xenoviruses. Platelet lysate (PL) can be considered as a suitable alternative containing a high level of growth factors and a low production cost. Three-step culture media supplemented with PL or fetal bovine serum (FBS) were used for proliferation and differentiation of CD34 umbilical cord blood stem cells to erythrocytes in co-culture with bone marrow mesenchymal stem cells (BM-MSCs). The cells were cultivated for 15 days and cell proliferation and expansion were assessed using cell counts at different days. Erythroid differentiation genes, CD71 and glycophorin A expression levels were evaluated. Maximum hematopoietic stem cells (HSCs) proliferation was observed on day 15 in PL-containing medium (99±17×10-fold). Gene expression and surface markers showed higher differentiation of cells in PL-containing medium. The results of this study indicate that PL can enhance erythroid proliferation and differentiation of CD34 HSCs. PL can also be used as a proper alternative for FBS in the culture medium and HSCs differentiation.
红细胞(RBC)输血是外科护理、创伤和贫血治疗中一种常用的支持性治疗方法。然而,由于血液资源有限、疾病传播可能性、免疫反应以及稀有血型的存在,体内红细胞生成似乎是输血的合适替代方法。细胞培养需要无血清或添加了极其昂贵动物血清的培养基,而动物血清可能会传播异种病毒。血小板裂解液(PL)可被视为一种合适的替代物,其含有高水平的生长因子且生产成本低。添加了PL或胎牛血清(FBS)的三步培养基用于与骨髓间充质干细胞(BM-MSCs)共培养时CD34脐带血干细胞向红细胞的增殖和分化。细胞培养15天,并在不同天数通过细胞计数评估细胞增殖和扩增情况。评估红系分化基因、CD71和血型糖蛋白A的表达水平。在含PL培养基中第15天观察到最大造血干细胞(HSCs)增殖(99±17×10倍)。基因表达和表面标志物显示含PL培养基中的细胞分化程度更高。本研究结果表明,PL可增强CD34 HSCs红系增殖和分化。PL还可作为培养基中FBS以及HSCs分化的合适替代物。