Lemons R, Forster S, Thoene J
Department of Pediatrics, University of Michigan, School of Medicine, Ann Arbor 48109.
Anal Biochem. 1988 Jul;172(1):219-27. doi: 10.1016/0003-2697(88)90435-6.
Exposure of cultured diploid fibroblasts to protease solutions induces a hyperpermeable state which permits entry of exogenous macromolecules directly into the cytosol. We have exploited this finding to devise a microinjection method whose chief advantages are simplicity and good retention of cell viability. Proteins successfully injected by this technique range from insulin to thyroglobulin. The amounts injected range from 4 X 10(5) to 5 X 10(6) molecules/cell.
将培养的二倍体成纤维细胞暴露于蛋白酶溶液中会诱导一种高通透性状态,这种状态允许外源性大分子直接进入细胞质。我们利用这一发现设计了一种显微注射方法,其主要优点是操作简单且细胞活力保持良好。通过该技术成功注射的蛋白质范围从胰岛素到甲状腺球蛋白。注射量范围为每细胞4×10⁵至5×10⁶个分子。