少突胶质细胞体外培养纯化及鉴定、细胞体外缺氧模型建立
发布时间：2019-01-02 08:03
【摘要】： 【目的】 1、观察体外培养少突胶质细胞的增殖和分化,观察其生物学特性;探讨获得细胞纯度较高的实验方法,为少突胶质细胞损伤后髓鞘形成障碍或脱髓鞘疾病的研究奠定基础。 2、建立体外少突胶质细胞缺氧培养模型。 【方法】 1、取新生SD大鼠脑皮质进行体外培养,根据星形胶质细胞和少突胶质细胞生长时间差异、细胞生长方式及细胞对培养层粘附等特性的不同,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判断纯度。 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。
[Abstract]:[objective] 1. To observe the proliferation and differentiation of oligodendrocytes cultured in vitro, and to observe the biological characteristics of oligodendrocytes. To explore the experimental method of obtaining high purity of cells, which will lay a foundation for the study of myelin formation disorder or demyelinating disease after oligodendrocyte injury. 2. The model of hypoxia culture of oligodendrocytes in vitro was established. [methods] 1. The cerebral cortex of neonatal SD rats was cultured in vitro. According to the difference of growth time between astrocytes and oligodendrocytes, the cell growth pattern and cell adhesion to culture layer were different. The cell types were identified and the purity was judged by inverted microscope and immunofluorescence staining. 2. Anoxia combined with low glucose culture was used to establish anoxic culture model in vitro. Morphologic changes of cells were observed by phase contrast microscope. [results] 1. High purity rat oligodendrocytes were obtained. Oligodendrocyte progenitor cells were A2B5 positive and mature oligodendrocytes were galactosinolide positive. 2. The cell survival rate decreased significantly after hypoxia for 6 hours, and the number of living cells decreased and the number of apoptotic cells increased with the prolongation of hypoxia time, and most of the cells were broken down by hypoxia for 10 hours. [conclusion] 1. The high purity rat oligodendrocytes can be obtained by twice oscillatory isolation and purification in constant temperature shaking bed and in conditioned medium. 2. Hypoglycemia combined with hypoxia culture model can establish a feasible model of oligodendrocyte hypoxia in vitro.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R-332
本文编号：2398236
[Abstract]:[objective] 1. To observe the proliferation and differentiation of oligodendrocytes cultured in vitro, and to observe the biological characteristics of oligodendrocytes. To explore the experimental method of obtaining high purity of cells, which will lay a foundation for the study of myelin formation disorder or demyelinating disease after oligodendrocyte injury. 2. The model of hypoxia culture of oligodendrocytes in vitro was established. [methods] 1. The cerebral cortex of neonatal SD rats was cultured in vitro. According to the difference of growth time between astrocytes and oligodendrocytes, the cell growth pattern and cell adhesion to culture layer were different. The cell types were identified and the purity was judged by inverted microscope and immunofluorescence staining. 2. Anoxia combined with low glucose culture was used to establish anoxic culture model in vitro. Morphologic changes of cells were observed by phase contrast microscope. [results] 1. High purity rat oligodendrocytes were obtained. Oligodendrocyte progenitor cells were A2B5 positive and mature oligodendrocytes were galactosinolide positive. 2. The cell survival rate decreased significantly after hypoxia for 6 hours, and the number of living cells decreased and the number of apoptotic cells increased with the prolongation of hypoxia time, and most of the cells were broken down by hypoxia for 10 hours. [conclusion] 1. The high purity rat oligodendrocytes can be obtained by twice oscillatory isolation and purification in constant temperature shaking bed and in conditioned medium. 2. Hypoglycemia combined with hypoxia culture model can establish a feasible model of oligodendrocyte hypoxia in vitro.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R-332
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相关期刊论文 前3条
1 伍亚民,马海涵,廖维宏;大鼠星形胶质细胞和少突胶质细胞的纯化培养与鉴定[J];创伤外科杂志;2000年04期
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