星形胶质细胞瘤干细胞的体外分离、培养与鉴定

In vitro isolation, culture and identification of brain tumor stem cells from human astrogliomas

背景:肿瘤干细胞理论认为肿瘤中存在一小部分具有无限增殖潜能和自我更新能力,能够分化为成熟细胞表型的干细胞样细胞,对肿瘤发生、增殖、侵袭起关键作用。目的:建立体外分离、培养与鉴定星形胶质细胞瘤干细胞的方法。方法:采用直接培养法分离培养星形胶质细胞瘤干细胞。参照神经干细胞培养条件,进行体外培养。观察其增殖、分化并进行巢蛋白、CD133免疫细胞化学鉴定和诱导分化后神经元特异性烯醇化酶、胶质纤维酸性蛋白及O4免疫细胞化学鉴定。结果与结论:培养7-10d,可形成大量悬浮生长巢蛋白及CD133免疫阳性的神经球,经诱导分化后细胞呈神经元特异性烯醇化酶、胶质纤维酸性蛋白或O4免疫阳性。提示星形胶质细胞瘤中存在具有神经干细胞特性的肿瘤干细胞。CD133和巢蛋白是星形胶质细胞瘤干细胞重要的表面标记,可以用于星形胶质细胞瘤干细胞的分离。

BACKGROUND： According to the theory of tumor stem cells, there are a small amount of stem cell-like cells that exhibit infinite proliferative potential and self-renewal capacity, can differentiate into cells with the phenotype of mature cells and play a key role in tumor production, proliferation and invasion. OBJECTIVE： To investigate the feasibility of isolation, culture and identification of brain tumor stem cells from human astrogliomas.METHODS： Brain tumor stem cells were isolated by primary culture from human astrogliomas. These cells were cultured under the culture condition of neural stem cells. The clone spheres were identified with immunocytochemistery for nestin and CD133. At the same time, differentiated cells were identified by immunocytochemistery for neuron specific enolase, glial fibrillary acidic protein and O4, respectively. RESULTS AND CONCLUSION： After 7-10 days of culture, a great number of neurospheres immunoreactive for nestin and CD133 were observed. After induced differentiation, these cells were immunoreactive for neuron specific enolase, glial fibrillary acidic protein and O4. These findings suggest that there are brain tumor stem cells with the characteristics of neural stem cells in human astrogliomas. CD133 and nestin are key surface markers for brain tumor stem cells, which can be used for isolation of brain tumor stem cells.