人眼视神经乳头星形胶质细胞体外培养:Ⅰ.原代培养和传代试验(英文)

Human optic nerve head astrocytes culture in vitro:Ⅰ. the primary culture and passage

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。

AIM： To culture astrocytes from human donor eyes in order to understand the function of astrocytes in remodelling events in the glaucomatous optic nerve head （ONH）. METHODS： Primary cultures were prepared by explantation of human ONH tissue in order to get astrocytes. Laminar criborsa （LC） cells were prepared concurrently for comparison. Astrocyte cultures could be separated from LC cells by selecting medium. Similar procedures were used for LC. RESULTS： Primary cells grew from human optic nerve head explants 4-8 weeks after explantation. Astrocytes had different morphologies and growth characteristics from LC cells. Type 1B astrocyte cells could grow in medium without FBS. Purified cultures were obtained by second passage and could be harvested by third to fifth passage, which were prepared to use for further study, including being characterized by positive glial fibrillary acidic protein （GFAP） and neural cell adhesion molecule （NCAM） staining. CONCLUSION： Precise dissection of fragment is the most important step to get clear explants for primary culture. Economic and rapid method could be useful to select cells by different mediums, which will help us to get more purified cells for further study.