Harman A W, McCamish L E, Henry C A
J Pharmacol Methods. 1987 Apr;17(2):157-63. doi: 10.1016/0160-5402(87)90026-x.
A method for the isolation of hepatocytes from postnatal (1- to 3-week-old) mice has been developed. Cell isolation was carried out by retrograde perfusion of the liver with a collagenase-containing bicarbonate buffer. Viable cells were separated by selective adsorption onto collagen membranes. Cell viability was assessed by measuring ATP and glutathione content, lactate:pyruvate ratio, and the rate of protein synthesis. Comparisons of these parameters were made with those in cells isolated from adult mice and with values in whole liver. These hepatocytes were capable of metabolizing acetaminophen to its known hepatic conjugates and were susceptible to acetaminophen toxicity. This procedure for isolating mouse hepatocytes should be useful for the study of hepatic drug metabolism and its relationship to toxicity in the postnatal mouse.
已开发出一种从出生后（1至3周龄）小鼠分离肝细胞的方法。通过用含胶原酶的碳酸氢盐缓冲液逆行灌注肝脏来进行细胞分离。通过选择性吸附到胶原膜上分离活细胞。通过测量ATP和谷胱甘肽含量、乳酸：丙酮酸比率以及蛋白质合成速率来评估细胞活力。将这些参数与从成年小鼠分离的细胞中的参数以及全肝中的值进行比较。这些肝细胞能够将对乙酰氨基酚代谢为其已知的肝脏共轭物，并且易受对乙酰氨基酚毒性的影响。这种分离小鼠肝细胞的方法对于研究出生后小鼠的肝脏药物代谢及其与毒性的关系应该是有用的。
