【摘要】： 【目的】 1、觀察體外培養(yǎng)少突膠質(zhì)細(xì)胞的增殖和分化,觀察其生物學(xué)特性;探討獲得細(xì)胞純度較高的實(shí)驗(yàn)方法,為少突膠質(zhì)細(xì)胞損傷后髓鞘形成障礙或脫髓鞘疾病的研究奠定基礎(chǔ)。 2、建立體外少突膠質(zhì)細(xì)胞缺氧培養(yǎng)模型。 【方法】 1、取新生SD大鼠腦皮質(zhì)進(jìn)行體外培養(yǎng),根據(jù)星形膠質(zhì)細(xì)胞和少突膠質(zhì)細(xì)胞生長(zhǎng)時(shí)間差異、細(xì)胞生長(zhǎng)方式及細(xì)胞對(duì)培養(yǎng)層粘附等特性的不同,采用兩次恒溫?fù)u床振蕩分離純化法和條件限定培養(yǎng)基培養(yǎng),倒置顯微鏡結(jié)合免疫熒光染色法鑒定細(xì)胞種類并判斷純度。 2、缺氧聯(lián)合低糖培養(yǎng)建立細(xì)胞體外缺氧培養(yǎng)模型,相差顯微鏡觀察細(xì)胞的形態(tài)學(xué)變化。 【結(jié)果】 1、獲取高純度的大鼠少突膠質(zhì)細(xì)胞。少突膠質(zhì)前體細(xì)胞祖細(xì)胞A2B5陽(yáng)性,成熟少突膠質(zhì)細(xì)胞半乳糖腦苷脂陽(yáng)性。 2、缺氧培養(yǎng)2小時(shí)細(xì)胞變化不明顯,但缺氧培養(yǎng)6小時(shí),細(xì)胞存活率明顯下降,并且隨著缺氧時(shí)間延長(zhǎng)存活細(xì)胞更少,凋亡細(xì)胞更多,至缺氧培養(yǎng)10小時(shí),大多數(shù)細(xì)胞破碎。 【結(jié)論】 1、兩次恒溫?fù)u床振蕩分離純化法和條件限定培養(yǎng)基培養(yǎng)可獲取高純度的大鼠少突膠質(zhì)細(xì)胞。 2、低糖聯(lián)合缺氧培養(yǎng)模型可以建立可行的少突膠質(zhì)細(xì)胞體外缺氧模型。
[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.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R-332

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本文編號(hào)：2398236