INTRODUCTION OF MACROMOLECULES INTO CULTURED MAMMALIAN-CELLS BY OSMOTIC LYSIS OF PINOCYTIC VESICLES
INTRODUCTION OF MACROMOLECULES INTO CULTURED MAMMALIAN-CELLS BY OSMOTIC LYSIS OF PINOCYTIC VESICLES
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DOI：
10.1016/0092-8674(82)90087-3
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发表时间：
1982-01-01
期刊：
影响因子：
64.5
通讯作者：
RECHSTEINER, M
中科院分区：
文献类型：
--
作者：
OKADA, CY;RECHSTEINER, M
基于胞饮小泡渗透裂解的哺乳动物细胞大分子导入新方法。首先将细胞在含有0.5M蔗糖、10%聚乙二醇1000和待转移的大分子的培养基中孵育。然后将细胞置于用0.66份水稀释的培养基中。大多数胞饮小泡在蔗糖存在下形成，在低渗介质中破裂，从而释放出封闭的大分子。L929细胞[小鼠胚胎成纤维细胞]在单次高渗蔗糖处理后保持完全存活，并且大多数在连续4轮渗透溶解中存活。该程序用于将大量辣根过氧化物酶、抗蓖麻毒素抗体和葡聚糖70，000转移到L929细胞的胞质溶胶中。直接比较由抗蓖麻毒素抗体转移所赋予的蓖麻毒素抗性程度，发现胞饮小体溶解与红细胞介导的注射相等，如果不是上级的话。
A new procedure for introducing macromolecules into cultured mammalian cells based on osmotic lysis of pinocytic vesicles. Cells are first incubated in culture medium containing 0.5 M sucrose, 10% polyethylene glycol 1000 and the macromolecule to be transferred. Cells are then placed in medium diluted with 0.66 parts water. Most pinocytic vesicles formed in the presence of sucrose burst in hypotonic medium, thereby releasing the enclosed macromolecule. L929 cells [mouse embryonic fibroblast] remain fully viable after a single hypertonic sucrose treatment, and a majority survives 4 successive rounds of osmotic lysis. This procedure used to transfer substantial amounts of horseradish peroxidase, antiricin antibodies and dextran 70,000 into the cytosol of L929 cells. Direct comparison of the degree of ricin resistance conferred by transfer of antiricin antibodies revealed pinosome lysis to be equal, if not superior, to injection mediated by red blood cells.