Mohammad-Gharibani Payam, Tiraihi Taki, Arabkheradmand Jalil
Dept of Anatomical Sciences, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Iran Neuroscience Center, Khatam-Al-Anbia Hospital, Tehran, Iran.
Iran Biomed J. 2009 Jul;13(3):137-43.
Cell therapy of many neurodegenerative diseases using bone marrow stromal cells (BMSC) requires the differentiation of BMSC into neuronal subtype. However, the transdifferentiation of BMSC into GABAergic phenotype requires more investigation.
In this study, BMSC of adult female rats were pre-induced into neuroblast-like cells using 1 mM beta-mercaptoethanol (betaME) and 10 microM retinoic acid (RA), followed by 40 mM potassium chloride as inducer. The BMSC were evaluated by fibronectin as well as Oct-4. The percentage of nestin, neurofilaments (NF 68, NF 160, and NF 200) and GABA immuno-reactive cells was used to evaluate the GABAergic differentiation at the pre-induction and induction stages. The statistical analysis was carried out using unpaired student's t-test and ANOVA with Tukey's multiple comparison.
The BMSC in the fourth passage expressed fibronectin up to 91.24 +/- 0.82%. The pre-induced cells after 2 days of RA exposure showed the expression of neuroblastic markers of nestin and NF68 (81.56 +/- 2.64% and 82.12 +/- 2.65%, respectively). The yield of GABAergic neurons with beta-ME for 1 h and RA as pre-inducer for 2 days followed by potassium chloride as inducer (40 mM for 3 days) was 60.64% +/- 1.97%. In addition, NF160 and NF200 were detected in the transdifferentiated cells. RT-PCR showed no expression of Oct-4 after the induction and pre-induction stages.
GABAergic-like neurons obtained from BMSC can be potentially used in cell transplanting for some neurodegenerative disorders.
使用骨髓基质细胞（BMSC）对许多神经退行性疾病进行细胞治疗需要将BMSC分化为神经元亚型。然而，BMSC向γ-氨基丁酸能（GABAergic）表型的转分化需要更多研究。
在本研究中，成年雌性大鼠的BMSC先用1 mMβ-巯基乙醇（βME）和10 μM视黄酸（RA）预诱导为神经母细胞样细胞，然后用40 mM氯化钾作为诱导剂。通过纤连蛋白以及八聚体结合转录因子4（Oct-4）对BMSC进行评估。在预诱导和诱导阶段，使用巢蛋白、神经丝（NF 68、NF 160和NF 200）和GABA免疫反应性细胞的百分比来评估GABA能分化。使用非配对学生t检验和带有Tukey多重比较的方差分析进行统计分析。
第四代BMSC表达纤连蛋白的比例高达91.24±0.82%。RA处理2天后的预诱导细胞显示巢蛋白和NF68的神经母细胞标志物表达（分别为81.56±2.64%和82.12±2.65%）。以β-ME处理1小时、RA作为预诱导剂处理2天，然后以氯化钾作为诱导剂（40 mM处理3天），GABA能神经元的产率为60.64%±1.97%。此外，在转分化细胞中检测到NF160和NF200。逆转录聚合酶链反应（RT-PCR）显示在诱导和预诱导阶段后Oct-4无表达。
从BMSC获得的GABA能样神经元可能可用于某些神经退行性疾病的细胞移植。
