Department of Urology, Zhongnan Hospital, Wuhan University, Wuhan, 430071, China.
Cell Mol Neurobiol. 2012 Nov;32(8):1255-63. doi: 10.1007/s10571-012-9850-2. Epub 2012 May 9.
It is reported that adipose-derived stem cells (ADSCs) had multilineage differentiation potential, and could differentiate into neuron-like cells induced by special induction media, which may provide a new idea for restoration of erectile dysfunction (ED) after cavernous nerve injury. The aim of this research was to explore the neuronal differentiation potential of ADSCs in vitro. ADSCs isolated from inguinal adipose tissue of rat were characterized by flow cytometry, and results showed that ADSCs were positive for mesenchymal stem cell markers CD90 and CD44, but negative for hematopoietic stem cell markers. ADSCs maintained self-renewing capacity and could differentiate into adipocytes and neurocytes under special culture condition. In this research, two methods were used to induce ADSCs. In method 1, ADSCs were treated with the preinduction medium including epithelium growth factor, basic fibroblast growth factor, and brain derived neurotrophic factor (BDNF) for 3 days, then with the neurogenic induction medium containing isobutylmethylxanthine, indomethacin, and insulin. While in method 2, BDNF was not used to treat ADSCs. After induction, neuronal differentiation of ADSCs was evaluated. Neuronal markers, glial fibrillary acidic protein (GFAP), and β-tubulin III (Tuj-1) were detected by immunofluorescence and Western Blot analyses. The expressions of GFAP and Tuj-1 in method 1 were obviously higher then those in method 2. In addition, the positive rate of the neuron-like cells was higher in method 1. It suggested that ADSCs are able to differentiate into neural-like cells in vitro, and the administration of BDNF in the preinduction medium may provide a new way to modify the culture method for getting more neuron-like cells in vitro.
据报道，脂肪来源的干细胞（ADSCs）具有多能分化潜能，可在特殊诱导培养基的诱导下分化为神经元样细胞，这可能为海绵体神经损伤后勃起功能障碍（ED）的恢复提供新的思路。本研究旨在探讨 ADSC 体外向神经元分化的潜能。从大鼠腹股沟脂肪组织中分离 ADSC，采用流式细胞术进行鉴定，结果显示 ADSC 表达间充质干细胞标志物 CD90 和 CD44，而不表达造血干细胞标志物。ADSC 具有自我更新能力，并可在特定培养条件下分化为脂肪细胞和神经细胞。本研究采用两种方法诱导 ADSC 向神经元分化。方法 1：用含表皮生长因子、碱性成纤维细胞生长因子和脑源性神经营养因子（BDNF）的预诱导培养基处理 ADSC 3 天，然后用含异丁基甲基黄嘌呤、吲哚美辛和胰岛素的神经诱导培养基处理。方法 2：BDNF 不用于处理 ADSC。诱导后，通过免疫荧光和 Western blot 分析检测神经元标志物胶质纤维酸性蛋白（GFAP）和β-微管蛋白 III（Tuj-1）的表达。结果显示，方法 1 中 GFAP 和 Tuj-1 的表达明显高于方法 2。此外，方法 1 中神经元样细胞的阳性率较高。这表明 ADSC 可在体外分化为神经元样细胞，BDNF 在预诱导培养基中的添加可能为改良体外获得更多神经元样细胞的培养方法提供新途径。
