Chen Jian-rong, Xu Ru-xiang, Jiang Xiao-dan, Xu Qiang, Cai Ying-qian, Zou Yu-xi, Ding Lian-shu
Institute of Neuromedicine of PLA, Zhujiang Hospital, First Military Medical University, Guangzhou 510280, China.
Di Yi Jun Yi Da Xue Xue Bao. 2004 Apr;24(4):367-70.
To study whether the neuron-like cells derived from bone marrow stromal cells (BMSCs) may excrete amino acids with neurobiological activities and possess the biochemical characteristics of neurons.
Under sterile condition, BMSCs from New Zealand rabbits were purified by gradient density centrifugation, and were induced to differentiate into neural stem cells and neuronal-like cells in the culture medium for neural stem cells containing retinoic acid (RA, 0.5 microg/ml) and glial-derived neurotrophic factor (GDNF, 20 ng/ml). The differentiated cells were then examined with immunocytochemical method and high-performance liquid chromatograpy (HPLC).
The round and enlarged BMSCs on day 10 of cell culture were positive for nestin, and on day 20, the cells with RA+GDNF stimulation differentiated into neuron-like cells with long protrusions and presented neuron-specific enolase (NSE) antigen. HPLC identified high levels of amino acids like Asp, Glu, Gly and Ala in the differentiated cells (P<0.01).
Rabbit BMSCs may proliferate in vitro as from nestin-positive cells and differentiate into NSE-positive cells containing high levels of excitatory and inhibitory amino acid neurotransmitters. RA and GDNF are important promoters for in vitro differentiation of the BMSCs toward neural stem cells.
研究源自骨髓基质细胞（BMSCs）的神经元样细胞是否可分泌具有神经生物学活性的氨基酸并具备神经元的生化特性。
在无菌条件下，采用梯度密度离心法纯化新西兰兔的骨髓基质细胞，并将其置于含有视黄酸（RA，0.5微克/毫升）和胶质细胞源性神经营养因子（GDNF，20纳克/毫升）的神经干细胞培养基中诱导分化为神经干细胞和神经元样细胞。然后采用免疫细胞化学方法和高效液相色谱法（HPLC）对分化后的细胞进行检测。
细胞培养第10天时，圆形且增大的骨髓基质细胞巢蛋白呈阳性，第20天时，经RA+GDNF刺激的细胞分化为具有长突起的神经元样细胞，并呈现神经元特异性烯醇化酶（NSE）抗原。HPLC鉴定出分化后的细胞中天门冬氨酸、谷氨酸、甘氨酸和丙氨酸等氨基酸含量较高（P<0.01）。
兔骨髓基质细胞可作为巢蛋白阳性细胞在体外增殖，并分化为含有高水平兴奋性和抑制性氨基酸神经递质的NSE阳性细胞。RA和GDNF是骨髓基质细胞体外向神经干细胞分化的重要促进因子。
