In vitro differentiation of size-sieved stem cells into electrically active neural cells
In vitro differentiation of size-sieved stem cells into electrically active neural cells
复制标题
DOI：
10.1634/stemcells.20-6-522
复制
发表时间：
2002-01-01
期刊：
影响因子：
5.2
通讯作者：
Ma, HL
中科院分区：
文献类型：
--
作者：
Hung, SC;Cheng, H;Ma, HL
从人骨髓中分离并在体外繁殖的尺寸筛选干细胞 (SS) 是具有一致标记分型的细胞群，可以形成骨骼、脂肪和软骨。在本实验中，我们证明了在实验细胞培养条件下可以诱导SS细胞分化为神经细胞。在无血清条件下暴露于抗氧化剂（β-巯基乙醇+/-视黄酸）五小时后，SS细胞表达巢蛋白、神经元特异性烯醇化酶（NSE）、神经元特异性核蛋白（NeuN）和神经元特异性微管蛋白-1（TuJ-1）的蛋白，以及NSE和Tau的mRNA。免疫荧光显示几乎所有细胞（>98%）均表达NeuN和TuJ-1。 β-巯基乙醇处理5天后，SS细胞表达神经丝高蛋白，但不表达丝裂原激活蛋白2、神经胶质丝酸性蛋白和半乳脑苷脂。对于这种长期处理的细胞，可以通过电生理记录检测电压敏感离子电流，并且可以通过高钾（K+）缓冲液和谷氨酸提高细胞内钙离子Ca2+浓度。这些发现表明，SS 细胞可能是神经功能障碍细胞治疗和基因治疗的未分化细胞的替代来源。
Size-sieved stem (SS) cells isolated from human bone marrow and propagated in vitro are a population of cells with consistent marker typing, and can form bone, fat, and cartilage. In this experiment, we demonstrated that SS cells could be induced to differentiate into neural cells under experimental cell culture conditions. Five hours after exposure to antioxidant agents (beta-mercaptoethanol +/- retinoic acid) in serum-free conditions, SS cells expressed the protein for nestin, neuron-specific enolase (NSE), neuron-specific nuclear protein (NeuN), and neuron-specific tubulin-1 (TuJ-1), and the mRNA for NSE and Tau. Immunofluorescence showed that almost all the cells (>98%) expressed NeuN and TuJ-1. After 5 days of beta-mercaptoethanol treatment, the SS cells expressed neurofilament high protein but not mitogen-activated protein-2, glial filament acidic protein, and galactocerebroside. For such long-term-treated cells, voltage-sensitive ionic current could be detected by electrophysiological recording, and the intracellular calcium ion, Ca2+, concentration can be elevated by high potassium (K+) buffer and glutamate. These findings suggest that SS cells may be an alternative source of undifferentiated cells for cell therapy and gene therapy in neural dysfunction.