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目的:探索人眼视神经乳头星形胶质细胞体外培养的方法,为进一步研究星形胶质细胞在青光眼性视神经病变中的作用打下基础。方法:取材新鲜人眼视神经乳头组织和筛板组织,进行星形胶质细胞和筛板细胞的体外培养与传代试验。结果:组织块培养4~8wk后,原代细胞开始生长,星形胶质细胞在形态学和生长特性上与筛板细胞明显不同,β1型星形胶质细胞可以在无血清的培养液中生长良好,通过无血清培养液选择性培养可以在第二代传代过程中分离出纯化的星形胶质细胞,并可在第三至第四代收获大量细胞以备后续的研究。结论:精细准确的组织解剖分离对于获得纯化细胞至关重要,采用无血清培养液选择分离获得星形胶质细胞方法经济简单,细胞纯度高,便于进一步储存和研究。
OBJECTIVE: To explore the method of in vitro culture of human optic nerve head astrocytes, and to lay a foundation for the further study of the role of astrocytes in glaucomatous optic neuropathy. Methods: Fresh human optic nerve head tissue and sieve plate were taken for in vitro culture and passage test of astrocytes and sieve plate cells. RESULTS: Primary cultured cells began to grow after 4 ~ 8 weeks of culture. Astrocytes obviously differed in morphology and growth characteristics from those of sieved plate, and β1-type astrocytes could be cultured in serum-free medium Well-grown, purified astrocytes can be isolated during second passage through selective cultures of serum-free medium and large numbers of cells can be harvested from the third to fourth generation for subsequent studies. Conclusion: The precise and accurate tissue dissection is essential for obtaining purified cells. The method of selecting and separating astrocytes with serum-free culture medium is simple and economical, and has high cell purity for further storage and study.