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An improved method for primary culture of rat podocytes A gentle method to isolate glomeruli simply by cutting renal cortices without forced sieving was devised in our previous study of primary podocyte culture.Yields of glomeruli isolated by this method, however, were too small to perform subculture or biological assays. In the present study, we tried an isolation method with magnetic beads and collagenase to increase the yields.Rat kidneys were perfused with magnetic particles.Renal cortices were digested with collagenase and filtered.Utilizing magnetic particles trapped within glomeruli,glomeruli were collected by attractive power of a magnet and cultured.The number of glomeruli isolated from one adult rat was more than 20000 and the purity was more than 97%. About half of them were attached to culture dishes and exhibited cellular outgrowths, which were identified as podocytes by their distinct staining for podocyte markers.After 3 days of primary culture, the cellular outgrowths were subcultured.Approximately 60 podocytes were obtained per attached glomerulus.Their significant expression of podocytes markers was demonstrated by immunostaining and quantitative reverse transcriptase-polymerase chain reaction. The isolation method with magnetic beads and collagenase provides a number of glomeruli suitable for primary podocyte culture. 普遍采用的一种分离肾小球的方法：简单的切取肾皮质而不进行过筛。然而，通过这种方法收集的肾小球数量太少，以至于不能进行传达培养和生物学实验。在最近的研究中，我们尝试了一种加入磁珠和胶原酶的分离方法来增加肾小球的产量。将磁珠灌满大鼠肾脏。胶原酶消化肾皮质并过滤。磁珠捕获肾小球，并利用磁铁的吸引力吸引肾小球进行培养。利用这种方法从一只成年大鼠皮质内可分离出超过20000个肾小球，并且纯度超过97%，其中大约一半会贴壁并生成副产物，通过对足细胞特异分子的染色鉴定为足细胞。原代培养3天后，进行传代培养。每个贴壁的肾小球可得到大约60个足细胞。通过免疫染色和定量逆转录聚合酶链反应证实足细胞标记物的显著表达。运用磁珠和胶原酶分离肾小球的这种方法可提供适合于足细胞原代培养的肾小球量。 Cell culture has been used to elucidate the physiological and pathophysiological characteristics of podocytes.Recently,there has been a tremendous increase in studies dealing with cultured podocytes,to which immortalized mouse1,2and human3 podocyte cell lines have made great contributions.However