Mu M W, Zhao Z Y, Li C G
Department of Orthopedics, Dongzhimen Hospital Beijing University of Chinese Medicine, Beijing, China.
Department of Orthopedics, Dongzhimen Hospital Beijing University of Chinese Medicine, Beijing, China
Genet Mol Res. 2015 Oct 30;14(4):14169-76. doi: 10.4238/2015.October.29.39.
Neurogenic differentiation of bone marrow (BM) mesenchymal stem cells (MSCs) offers a new hope for patients with many neurological disorders. Several chemical inducers are employed to induce BMMSCs differentiation into nerve cells. In the present study, we compared different inducers [2-mercaptoethanol (BME), tretinoin (ATRA), dimethyl sulfoxide/butylated hydroxyanisole (DMSO/BHA), and indomethacin/3-isobutyl-1-methylxanthine (indomethacin/IBMX)] on the neurogenic differentiation of BMMSCs and aimed to identify a more efficient and safer method. The MSCs were first identified by their ability to adhere to plastic and by the expression of positive (CD44, CD90, and CD105) and negative (CD34) markers assessed by flow cytometry. The efficiency of the neurogenic differentiation was determined by assessing the mRNA and protein expression of nestin, microtubule-associated protein-2 (MAP2), neuron specific enolase (NSE), and glial fibrillary acidic protein (GFAP) by reverse transcription-polymerase chain reaction and western-blot, respectively. The effect of these inducers on cell viability was also evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. This comprehensive study shows that indomethacin/IBMX is better than BME, DMSO/BHA, and ATRA both in terms of efficiency and safety, while BME suppressed the growth and proliferation of MSCs.
骨髓间充质干细胞（MSCs）的神经源性分化为许多神经系统疾病患者带来了新希望。几种化学诱导剂被用于诱导骨髓间充质干细胞分化为神经细胞。在本研究中，我们比较了不同诱导剂[2-巯基乙醇（BME）、维甲酸（ATRA）、二甲基亚砜/叔丁基对苯二酚（DMSO/BHA）和吲哚美辛/3-异丁基-1-甲基黄嘌呤（吲哚美辛/异丁基甲基黄嘌呤）]对骨髓间充质干细胞神经源性分化的影响，旨在确定一种更有效且更安全的方法。首先通过其贴壁能力以及通过流式细胞术评估的阳性（CD44、CD90和CD105）和阴性（CD34）标志物的表达来鉴定间充质干细胞。分别通过逆转录-聚合酶链反应和蛋白质免疫印迹法评估巢蛋白、微管相关蛋白-2（MAP2）、神经元特异性烯醇化酶（NSE）和胶质纤维酸性蛋白（GFAP）的mRNA和蛋白质表达，以确定神经源性分化的效率。还通过3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐试验评估这些诱导剂对细胞活力的影响。这项综合研究表明，吲哚美辛/异丁基甲基黄嘌呤在效率和安全性方面均优于BME、DMSO/BHA和ATRA，而BME抑制了间充质干细胞的生长和增殖。
