Ghorbanian Mohammad, Mirzaeian Leila, Ghorbanian Mohammad Taghi, Rostami Farzaneh
Department of Cellular and Molecular Biology, School of Biology, Damghan University, Damghan, Iran.
Department of Embryology, Reproductive Biomedicine Research Centre, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran.
Cell J. 2023 Jun 28;25(6):383-390. doi: 10.22074/cellj.2023.1987841.1214.
Neural stem cells (NSCs) are suitable therapeutic candidates. Here, we compare the proliferation rate, differentiation potential, and expression levels of specific markers in two groups of cultured NSCs derived from rat subgranular (SGZ) and subventricular (SVZ) zones.
In this experimental study, NSCs isolated from SGZ and SVZ were cultured in α-minimal essential medium (α-MEM) supplemented with 1% penicillin/streptomycin, 10% fetal bovine serum (FBS), 20 ng/ml basic fibroblast growth factor (bFGF), 20 ng/ml epidermal growth factor (EGF), and B27 supplement. Glial fibrillary acidic protein (), p75 neurotrophin receptor (), tyrosine kinase receptor A (), beta-tubulin III (), and Nestin gene levels were compared via reverse transcription polymerase chain reaction (RT-PCR) in these NSCs. Nestin and Gfap protein levels were compared by immunoassay. Subsequently, both populations were induced with 10-8 M selegiline for 48 hours, followed by immunohistochemical analysis of tyrosine hydroxylase (TH) levels. One-way ANOVA and Tukey's post-test were used with a significance level of P<0.05.
Both groups were successfully expanded and expressed the neurotrophin receptor genes. The SGZNSCs had a significantly higher proliferation rate and significantly higher numbers of Nestin and Gfap-positive cells. Although the majority of selegiline-induced NSCs were TH-positive, we observed more TH-positive cells in SGZ-derived NSCs and these SGZ-NSCs displayed a shorter differentiation time.
SGZ-derived NSCs appear to be a more appropriate candidate for therapeutic purposes based on proliferation rate, neurosphere size, and and expression levels, as well as differentiation time and TH expression level after dopaminergic induction.
神经干细胞（NSCs）是合适的治疗候选者。在此，我们比较了来自大鼠颗粒下区（SGZ）和脑室下区（SVZ）的两组培养神经干细胞的增殖率、分化潜能以及特定标志物的表达水平。
在本实验研究中，从SGZ和SVZ分离的神经干细胞在补充有1%青霉素/链霉素、10%胎牛血清（FBS）、20 ng/ml碱性成纤维细胞生长因子（bFGF）、20 ng/ml表皮生长因子（EGF）和B27添加剂的α-最低必需培养基（α-MEM）中培养。通过逆转录聚合酶链反应（RT-PCR）比较这些神经干细胞中胶质纤维酸性蛋白（）、p75神经营养因子受体（）、酪氨酸激酶受体A（）、β-微管蛋白III（）和巢蛋白基因水平。通过免疫测定比较巢蛋白和胶质纤维酸性蛋白的蛋白水平。随后，两组细胞均用10 - 8 M司来吉兰诱导48小时，然后对酪氨酸羟化酶（TH）水平进行免疫组织化学分析。采用单因素方差分析和Tukey事后检验，显著性水平为P<0.05。
两组均成功扩增并表达神经营养因子受体基因。SGZ神经干细胞的增殖率显著更高，巢蛋白和胶质纤维酸性蛋白阳性细胞数量也显著更多。尽管大多数司来吉兰诱导的神经干细胞为TH阳性，但我们在源自SGZ的神经干细胞中观察到更多TH阳性细胞，且这些SGZ神经干细胞的分化时间更短。
基于增殖率、神经球大小、以及和表达水平，以及多巴胺能诱导后的分化时间和TH表达水平，源自SGZ的神经干细胞似乎是更适合用于治疗目的的候选者。
