Cell J. 2014 Summer;16(2):195-202. Epub 2014 May 25.
Mesenchymal stem cells (MSCs) can differentiate into various cell types. Since cysteine has structural similarities to neuronal inducers β-mercaptoethanol and glutathione, we examined its effect on neural induction of rat bone marrow MSCs.
In this experimental study, cells were treated in a medium containing 1mM cysteine for 24 hours prior to treatment with neuron inducing medium containing 10 mM cysteine for 1, 2 and 3 hours. Cell viability and morphology were assessed by 3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide (MTT) assay and, Hoechst, propidium iodide and acridine orange staining respectively. Expression of nestin and β-Tubulin III genes, as neural cell-specific markers, was studied reverse transcription polymerase chain reaction (RT-PCR). The data was statistically analyzed using One-Way ANOVA and Tukey's test and p<0.05 was considered significant.
After 3 hours of treatment, neuron like morphology with a considerable expression of nestin and β-Tubulin III genes was apparent. The mean cell viability was not significantly different at 1, 2 and 3 hours following induction, compared with the control cells.
Cysteine can induce neural features in rat bone marrow MSCs without reducing cell viability. Therefore, it can be considered as a safer alternative to toxic neural inducer agents such as β-mercaptoethanol.
间充质干细胞(MSCs)可分化为多种细胞类型。由于半胱氨酸在结构上与神经元诱导剂β-巯基乙醇和谷胱甘肽相似,我们研究了其对大鼠骨髓间充质干细胞向神经诱导的影响。
在这项实验研究中,细胞在含有 1mM 半胱氨酸的培养基中处理 24 小时,然后用含有 10mM 半胱氨酸的神经元诱导培养基处理 1、2 和 3 小时。细胞活力和形态分别通过 3-(4,5-二甲基噻唑-2-Yl)-2,5-二苯基四唑溴盐(MTT)测定和 Hoechst、碘化丙啶和吖啶橙染色进行评估。巢蛋白和β-Tubulin III 基因的表达作为神经细胞特异性标志物,通过逆转录聚合酶链反应(RT-PCR)进行研究。使用单因素方差分析和 Tukey 检验对数据进行统计学分析,p<0.05 被认为具有统计学意义。
诱导 3 小时后,可见具有明显巢蛋白和β-Tubulin III 基因表达的神经元样形态。与对照细胞相比,诱导后 1、2 和 3 小时的细胞活力没有显著差异。
半胱氨酸可诱导大鼠骨髓间充质干细胞产生神经特征,而不会降低细胞活力。因此,它可以被认为是比β-巯基乙醇等有毒神经诱导剂更安全的替代品。