Chai Li-Hui, Wu Su-Xia, Yan Wen-Hai, Ma Yuan-Fang
Laboratory for Cell and Molecular Immunology of Henan University, Institute for Immunology of Henan University, Kaifeng 475004, China.
Sheng Wu Gong Cheng Xue Bao. 2007 Mar;23(2):252-6.
Midbrain dopamine (DA) neurons play an essential role in modulating motor control. Defects in central DA neurons affect a wide range of neurological disorders including Parkinson's disease (PD). The greatest motivation in the field has been the potential use of DA neurons for cell transplantation therapy in Parkinsonian patients. Recent studies indicated that BMSCs could differentiate into DA neurons in vitro as neural stem cells (NSC) and embryonic stem cells (ESC) could. However, there are no direct evidences about functional DA neurons derived from BMSCs. According to the protocols which had been applicated in inducing neuronal stem cells and embryonic stem cells differentiate into DA neurons in vitro, the present study provides a protocol by using 50 micromol/L brain derived neurotrophy factor (BDNF), 10 micromol/L forskolin (FSK) and 10 micromol/L dopamine (DA) to induce BMSCs differentiate into DA neurons. After 2 weeks of differentiation, the cells expressed the character of neurons in ultrastructure. RT-PCR discovered mRNA of NSE (neuron specific enolase), Nurr1, Ptx3, Lmx1b and Tyrosine hydroxylase (TH) were positive. Immunocytochemistry staining indicated the ratio of TH-positive neural cells was significantly increased after induced 2 weeks (24.80 +/- 3.36) % compared to that of induction of 3 days (3.77 +/- 1.77) %. And the DA release was also different between differentiated and undifferentiated cells detected by high performance liquid chromatography (HPLC). That is to say BDNF and FSK and DA can induce BMSCs differentiate into DA neurons in vitro, and the transdifferentiated cells express mature neurons characters. BMSCs might be a suitable and available source for the in vitro derivation of DA neurons and cell transplantation therapy in some central neural system diseases such as PD.
中脑多巴胺（DA）神经元在调节运动控制中起着至关重要的作用。中枢DA神经元的缺陷会影响包括帕金森病（PD）在内的多种神经系统疾病。该领域最大的动力在于DA神经元在帕金森病患者细胞移植治疗中的潜在应用。最近的研究表明，骨髓间充质干细胞（BMSCs）能够像神经干细胞（NSCs）和胚胎干细胞（ESCs）一样在体外分化为DA神经元。然而，目前尚无关于源自BMSCs的功能性DA神经元的直接证据。根据已应用于诱导神经干细胞和胚胎干细胞在体外分化为DA神经元的方案，本研究提供了一种使用50微摩尔/升脑源性神经营养因子（BDNF）、10微摩尔/升福斯可林（FSK）和10微摩尔/升多巴胺（DA）诱导BMSCs分化为DA神经元的方案。分化2周后，细胞在超微结构上表现出神经元的特征。逆转录-聚合酶链反应（RT-PCR）发现神经元特异性烯醇化酶（NSE）、核受体相关因子1（Nurr1）、垂体瘤转化基因3（Ptx3）、LIM同源框转录因子1β（Lmx1b）和酪氨酸羟化酶（TH）的信使核糖核酸（mRNA）呈阳性。免疫细胞化学染色表明，诱导2周后TH阳性神经细胞的比例（24.80±3.36）%显著高于诱导3天后的比例（3.77±1.77）%。通过高效液相色谱法（HPLC）检测，分化细胞和未分化细胞之间的DA释放也存在差异。也就是说，BDNF、FSK和DA能够在体外诱导BMSCs分化为DA神经元，且转分化细胞表现出成熟神经元的特征。BMSCs可能是体外获得DA神经元以及在某些中枢神经系统疾病如PD中进行细胞移植治疗的合适且可用的细胞来源。
