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司来吉兰诱导脂肪组织来源干细胞转分化为表达神经营养因子的神经元样细胞

Trans-differentiation of the adipose tissue-derived stem cells into neuron-like cells expressing neurotrophins by selegiline.

作者信息

Abdanipour Alireza, Tiraihi Taki, Delshad Alireza

机构信息

Dept. of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

出版信息

Iran Biomed J. 2011;15(4):113-21. doi: 10.6091/ibj.1011.2012.

Abstract

BACKGROUND

Adult stem cells (ASC) are undifferentiated cells found throughout the body. These cells are promising tools for cell replacement therapy in neurodegenerative disease. Adipose tissue is the most abundant and accessible source of ASC. This study was conducted to evaluate effect of selegiline on differentiation of adipose-derived stem cells (ADSC) into functional neuron-like cells (NLC), and also level of the neurotrophin expression in differentiated cells.

METHODS

ADSC were transdifferentiated into NLC using selegiline where CD90, CD49d, CD31, CD106 and CD45 were used as markers for ADSC identification. Lipogenic and osteogenic differentiation of ADSC were used to characterize the ADSC. ADSC were treated with selegiline at different concentrations (from 10(-6) to 10(-11) mM) and time points (3, 6, 12, 24 and 48 h). Percentage of viable cells, nestin and neurofilament 68 (NF-68) immunoreactive cells were used as markers for differentiation. The optimal dose for neurotrophin expressions in differentiating cells was evaluated using reverse transcriptase-PCR. NLC function was evaluated by loading and unloading with FM1-43 dye.

RESULTS

ADSC were immunoreactive to CD90 (95.67 ± 2.26), CD49d (71.52 ± 6.64) and CD31 (0.6 ± 0.86), but no immunoreactivity was detected for CD106 and CD45. The results of neural differentiation showed the highest percentage of nestin and NF-68 positive cells at 10(-9) mM concentration of selegiline (exposed for 24 h). The differentiated cells expressed synapsin and neurotrophin genes except brain-derived neurotrophic factor.

CONCLUSION

ADSC can be an alternative source in cell-based therapy for neurodegenerative diseases using selegiline to induce ADSC differentiation to neuronal lineage.

摘要

背景

成体干细胞(ASC)是遍布全身的未分化细胞。这些细胞是神经退行性疾病细胞替代疗法中很有前景的工具。脂肪组织是ASC最丰富且最易获取的来源。本研究旨在评估司来吉兰对脂肪来源干细胞(ADSC)向功能性神经元样细胞(NLC)分化的影响,以及分化细胞中神经营养因子的表达水平。

方法

使用司来吉兰将ADSC转分化为NLC,其中CD90、CD49d、CD31、CD106和CD45用作ADSC鉴定的标志物。ADSC的脂肪生成和成骨分化用于表征ADSC。用不同浓度(从10⁻⁶到10⁻¹¹ mM)和时间点(3、6、12、24和48小时)的司来吉兰处理ADSC。活细胞百分比、巢蛋白和神经丝68(NF - 68)免疫反应性细胞用作分化的标志物。使用逆转录聚合酶链反应评估分化细胞中神经营养因子表达的最佳剂量。通过用FM1 - 43染料加载和卸载来评估NLC功能。

结果

ADSC对CD90(95.67±2.26)、CD49d(71.52±6.64)和CD31(0.6±0.86)呈免疫反应性,但未检测到CD106和CD45的免疫反应性。神经分化结果显示,在司来吉兰浓度为10⁻⁹ mM(暴露24小时)时,巢蛋白和NF - 68阳性细胞的百分比最高。分化细胞表达突触素和神经营养因子基因,但不表达脑源性神经营养因子。

结论

ADSC可以成为神经退行性疾病基于细胞治疗的替代来源,使用司来吉兰诱导ADSC向神经元谱系分化。

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