Xu J, Lu H, Miao Z-N, Wu W-J, -Z Jiang Y, Ge F, Fang W-F, Zhu A-H, Chen G, Zhou J-H, Lu Y-Z, Tang Z-F, Wang Y
Wuxi Third People's Hospital, Liangxi, Wuxi, Jiangsu, China.
Eur Rev Med Pharmacol Sci. 2016 Dec;20(24):5041-5048.
To evaluate the immune activity of bone marrow mesenchymal stem cells (BMSCs), and explore the biological characteristics and capabilities of BMSCs and the potential to be differentiated into neuronal cells in vitro.
The BMSCs were isolated and proliferated in vitro to generate the xenogeneic mixed lymphocyte reaction. Moreover, peripheral BMSCs (pBMSCs) were added according to different ratios, which methods were stated as follows: 1: Dulbecco's Modified Eagle Medium (DMEM) + 10% Fetal Bovine Serum (FBS) + 1 μmol/L all-trans-retinoic acid (ATRA) + 20 μg/L basic fibroblast growth factor (bFGF) + 20 μg/L epidermal growth factor (EGF); 2: DMEM + 2% dimethyl sulfoxide (DMSO) + 100 μmol/L butylated hydroxyanisole (BHA). The immunofluorescence and immunohistochemical staining were finally used to evaluate the differentiation capabilities of human BMSCs (hBMSCs) induced in neuronal cells.
hBMSCs inhibited the lymphocyte proliferation in the mixed lymphocyte reaction (MLR) system at a proportional inhibition rate with additional numbers of stem cells. At hour 2 after culture with method 1, the plasma of hBMSCs shrank to nuclei and perinuclear bodies and was visualized under the light microscope. At hours 3-5, most of the hBMSCs formed neuron-like cells with total cell number unchanged. Afterward, the hBMSCs turned into bipolar or multipolar shaped cells and interconnected into a large network at Day 3. With immunofluorescence and immunohistochemical staining, 60-70% of the hBMSCs showed neurospecific enolase (NSE) positive and 45-50% glial fibrillary acidic protein (GFAP) positive while the Nestin-positive cells decreased to 3.4%. However, when cultured 2 hours with method 2, the most of the hBMSCs formed bipolar or multipolar shaped cells, then died after 48 hours. 40-50% NSE and 35-40% GFAP were positively expressed. Significantly, the rate of Nestin-positive cells decreased from 63% to 1.6% from hour 2 after culture to hour 48.
hBMSCs may be effective for cell therapy and tissue engineering for the capability of differentiating into neuronal-like cells, as well as the capability of inhibiting lymphocyte proliferation in MLR system.
评估骨髓间充质干细胞（BMSCs）的免疫活性，探讨BMSCs的生物学特性和能力以及其在体外分化为神经元细胞的潜力。
分离并体外扩增BMSCs以产生异种混合淋巴细胞反应。此外，按照不同比例添加外周血BMSCs（pBMSCs），方法如下：1：杜氏改良 Eagle 培养基（DMEM）+10%胎牛血清（FBS）+1μmol/L 全反式维甲酸（ATRA）+20μg/L碱性成纤维细胞生长因子（bFGF）+20μg/L表皮生长因子（EGF）；2：DMEM+2%二甲基亚砜（DMSO）+100μmol/L丁基羟基茴香醚（BHA）。最后采用免疫荧光和免疫组织化学染色评估人BMSCs（hBMSCs）向神经元细胞诱导的分化能力。
hBMSCs在混合淋巴细胞反应（MLR）系统中以与干细胞数量增加成比例的抑制率抑制淋巴细胞增殖。用方法1培养2小时后，hBMSCs的细胞质收缩至细胞核和核周体，在光学显微镜下可见。在3 - 5小时，大多数hBMSCs形成神经元样细胞，细胞总数不变。之后，hBMSCs在第3天变成双极或多极形细胞并相互连接成一个大网络。通过免疫荧光和免疫组织化学染色，60 - 70%的hBMSCs显示神经特异性烯醇化酶（NSE）阳性，45 - 50%胶质纤维酸性蛋白（GFAP）阳性，而巢蛋白阳性细胞降至3.4%。然而，用方法2培养2小时后，大多数hBMSCs形成双极或多极形细胞，然后在48小时后死亡。40 - 50%的NSE和35 - 40%的GFAP呈阳性表达。值得注意的是，从培养后2小时到48小时，巢蛋白阳性细胞率从63%降至1.6%。
hBMSCs可能对细胞治疗和组织工程有效，因为其具有分化为神经元样细胞的能力以及在MLR系统中抑制淋巴细胞增殖的能力。
