脂肪基质细胞诱导分化为神经元细胞的实验

Neuronal differentiation of adipose tissue-derived stromal cells

背景:寻找合适的种子细胞是移植治疗脑血管疾病及其他中枢神经系统疾病的关键。目的:观察人脂肪组织来源的基质细胞分化为神经元细胞的能力。方法:脂肪组织取自要求去除腹部多余脂肪的健康成人,供者无传染性疾病和内分泌疾病。分离培养脂肪组织来源的基质细胞,采用神经诱导培养基加GM1对其进行诱导培养。倒置相差显微镜下连续观察细胞生长情况和形态变化,免疫组织化学法鉴定神经前体细胞的特异性标志神经巢蛋白、神经元特异性烯醇化酶和微管联合蛋白2的表达情况。结果与结论:①经神经诱导培养基加GM1诱导后,分化后的细胞大部分呈典型的神经元样细胞形态。②倒置相差显微镜下可见,于诱导后1h出现神经巢蛋白表达阳性,5h出现神经元特异性烯醇化酶和微管联合蛋白2表达阳性。提示脂肪基质细胞可分化为神经元细胞,分化后的神经元细胞具有表达神经巢蛋白、神经元特异性烯醇化酶和微管联合蛋白2的功能。

BACKGROUND： Appropriate seed cell is important for transplantation in the treatment of cerebrovascular disease and other central nervous system disease. OBJECTIVE： To investigate the capacity of human adipose tissue-derived stromal cells （ADSCs） to differentiate into neurons. METHODS： The fatty tissue was harvested from removed abdominal unnecessary fat of healthy adult with no communicable disease or endocrine disease. Human ADSCs were isolated from human liposuction tissues and cultured in neural induction medium with GM1. Invert phase-contrast microscopy was used to observe morphology changes of ADSCs. The expression of nestin, neuron specific enolase （NSE） and microtubule-associated protein 2 （MAP2） were identified by immunocytochemistry. RESULTS AND CONCLUSION： The majority of cells displayed typical appearance of neuronal-like cells following induction. Following 1 hours of induction, some cells began to express nestin, and NSE and MAP2-positive cells were observed at 5 hours. ADSCs can differentiate into neurons, and the differentiated neurons have the capacity of expressing nestin, NSE and MAP2.