Pashoutan Sarvar Davod, Karimi Mohammad Hossein, Movassaghpour Aliakbar, Akbarzadehlaleh Parvin, Aqmasheh Sara, Timari Hamze, Shamsasenjan Karim
Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Transplant Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Adv Pharm Bull. 2018 Jun;8(2):291-296. doi: 10.15171/apb.2018.034. Epub 2018 Jun 19.
Mesenchymal stem cells (MSCs) play an important role in the proliferation and differentiation of hematopoietic stem cells (HSCs) in the bone marrow via cell-to-cell contact, as well as secretion of cytokines and microvesicles (MVs). In this study, we investigated the effect of mesenchymal stem cell-derived microvesicles (MSC-MVs) on erythroid differentiation of umbilical cord blood-derived CD34 cells. In this descriptive study, CD34 cells were cultured with mixture of SCF (10 ng/ml) and rhEPO (5 U/ml) cytokines in complete IMDM medium as positive control group. Then, in MV1- and MV2-groups, microvesicles at 10 and 20 µg/ml concentration were added. After 72 hours, erythroid specific markers (CD71 and CD235a) and genes (HBG1, GATA1, FOG1 and NFE2) were assessed by flow cytometry and qRT-PCR, respectively. The expression of specific markers of the erythroid lineages (CD71 and GPA) in the presence of different concentration of microvesicles were lower than that of the control group (P<0.001). Also, the expression of specific genes of the erythroid lineages (NFE2, FOG1, GATA1, and HBG1) was investigated in comparison to the internal control (GAPDH). Among all of them, HBG1 and FOG1 genes were significantly decreased to the control group (P<0.0001) but GATA1 and NFE2 gene expressions was not significant. The results of this study showed that MSC-MVs decrease the erythroid differentiation of umbilical cord blood-derived CD34 cells. Therefore, MSC-MVs play a key role in the regulation of normal erythropoiesis.
间充质干细胞（MSCs）通过细胞间接触以及细胞因子和微泡（MVs）的分泌，在骨髓中造血干细胞（HSCs）的增殖和分化中发挥重要作用。在本研究中，我们调查了间充质干细胞来源的微泡（MSC-MVs）对脐带血来源的CD34细胞红系分化的影响。在这项描述性研究中，将CD34细胞与SCF（10 ng/ml）和rhEPO（5 U/ml）细胞因子的混合物在完全IMDM培养基中培养作为阳性对照组。然后，在MV1组和MV2组中，分别添加浓度为10和20 µg/ml的微泡。72小时后，分别通过流式细胞术和qRT-PCR评估红系特异性标志物（CD71和CD235a）和基因（HBG1、GATA1、FOG1和NFE2）。在不同浓度微泡存在下，红系谱系特异性标志物（CD71和GPA）的表达低于对照组（P<0.001）。此外，与内参（GAPDH）相比，研究了红系谱系特异性基因（NFE2、FOG1、GATA1和HBG1）的表达。在所有这些基因中，HBG1和FOG1基因与对照组相比显著降低（P<0.0001），但GATA1和NFE2基因表达无显著差异。本研究结果表明，MSC-MVs降低了脐带血来源的CD34细胞的红系分化。因此，MSC-MVs在正常红细胞生成的调节中起关键作用。
