摘要
目的观察体外培养少突胶质细胞的增殖、分化及其生物学特性;探讨获得细胞纯度较高的实验方法及如何建立少突胶质细胞的缺氧培养模型。方法1.取新生SD大鼠脑皮质进行体外培养,采用两次恒温摇床振荡分离纯化法和条件限定培养基培养,倒置显微镜结合免疫荧光染色法鉴定细胞种类并分析纯度。2.缺氧联合低糖培养建立细胞体外缺氧培养模型,相差显微镜观察细胞的形态学变化。结果1.获取高纯度的大鼠少突胶质细胞。少突胶质前体细胞祖细胞A2B5荧光标记阳性,成熟少突胶质细胞半乳糖脑苷脂阳性。2.缺氧培养2小时细胞变化不明显,但缺氧培养6小时,细胞存活率明显下降,并且随着缺氧时间延长存活细胞更少,凋亡细胞更多,至缺氧培养10小时,大多数细胞破碎。结论1.两次恒温摇床振荡分离纯化法和条件限定培养基培养可获取高纯度的大鼠少突胶质细胞。2.低糖联合缺氧培养模型可以建立可行的少突胶质细胞体外缺氧模型。
Objective To culture, purify and identify the astroeytes and oligodendrocytes from rat cerebral tissue. And to study the influence of oxygen and glucose deprivation on oligodendrocyte type 2 astrocyte （O2A）progenitors. Methods Based on the different properties of cellular adhesions, developmental time-courses and growth pattern of astroglial cells and oligodendrocytes, astrocytes and oligodendrocytes were cultured and purified by twice orbital shaking, and identified by immmunofluorecence technique. O2A progenitors were cultured in oxygen deprivation combining with low glucose medium to mimic hypoxic injury. Results （1） Astrocytes and oligodendrocytes in mixed culture appeared better growth and differentiation than the respectively purified neuroglial, especially for the oligodendrocytes. O2A progenitors were A2BS-positive. The mature oligodendrocytes were Gal-positive. （2） Electron microscope showed a bundle of filament in the cytoplasma of O2A progenitors. The number of apoptotic O2A cells increased after oxygen deprivation combined with low glucose for 6 hours, and the apoptotic cells increased with time dependence. Most cells undergo necrosis after culturing in oxygen deprivation combined with low glucose medium for 10 hours. Conclusions （1） High ratios of purified oligodendrocytes can be obtained by twice orbital shaking and proper culture medium. （2） Oxygen deprivation combined with low glucose on O2A cultivation can mimic the ischemic-hypoxic changes on O2A progenitor in vivo.
出处
《四川解剖学杂志》
2009年第4期1-4,共4页
Sichuan Journal of Anatomy
