Ortiz D, Baldwin M M, Lucas J J
Department of Molecular and Cellular Biology, National Jewish Center for Immunology and Respiratory Medicine, Denver, Colorado 80206.
Mol Cell Biol. 1987 Aug;7(8):3012-7. doi: 10.1128/mcb.7.8.3012-3017.1987.
Material was introduced into cultures of cells by using the method of scrape loading, in which cells are simply rubbed from the surface of a plastic tissue culture dish by a rubber-tipped rod in the presence of a macromolecule of interest. The volume of solution introduced into cells was comparable to that generally injected in the direct microinjection method with glass capillaries, that is, about 50 to 100 fl per cell. Genetic defects (lack of hypoxanthine-guanine phosphoribosyltransferase and thymidine kinase) in several cell lines were transiently corrected by scraping the cells in the presence of crude cell extracts prepared from wild-type cells.
通过刮擦加载法将物质引入细胞培养物中,在该方法中,在存在感兴趣的大分子的情况下,用橡胶头棒简单地从塑料组织培养皿表面刮下细胞。引入细胞的溶液体积与通常用玻璃毛细管直接显微注射法注射的体积相当,即每个细胞约50至100飞升。通过在由野生型细胞制备的粗细胞提取物存在下刮擦细胞,几种细胞系中的遗传缺陷(次黄嘌呤 - 鸟嘌呤磷酸核糖转移酶和胸苷激酶缺乏)被短暂纠正。