小中大
Livers of 1-year-old male Wistar rats (450-500 g body mass, Interfauna), fed ad libitum on a stock diet, were perfused first with a Ca2+/Mg2+-free solution (137 mM NaCl, 5.4 mM KCl, 0.6 mM NaH2P04 . 2H20, 0.8 mM NaHPO4.12H20, 10 mM Hepes, 0.5 mM EGTA, 4.2 mM NaHCO3, and 5 mM glucose, pH 7.4) for 10 min at 37°C and next with 0.05% collagenase solution (137 mM NaC1, 5.4 mM KC1, 0.6mM NaH2PO4.2H20, 0.8mM Na2HPO4.12H20, 3.8mM CaCl2, 10mM Hepes, 4.2mM NaHCO3 and 500 mg/l collagenase, pH 7.4) for 30 min at 37°C. The flow rate was 10ml/min. The digested liver was excised, dispersed in Ca2+/Mg2+-free solution and filtered through gauzes. Residual hepatocytes were removed twice by a low-speed centrifugation (50 g, 4"C, 2 min). The non-parenchy-ma1 cells were pelleted by centrifugation (450 g, 4"C, 10 min). A stellate-cell-enriched fraction was obtained by the use of centrifugation with a triple-layered (9, 11, 17%) Nycodenz cushion (1400 g, 4"C, 20 min). The cells in the upper layer were washed by centrifugation (450g, 4"C, 10min) and suspended in DMEM (GIBCO) supplemented with 10% fetal-calf serum (Boehringer Mannheim) and antibiotics (105U/1 penicillin G, 100 mg/l streptomycin, and 2.5 mg/l amphotericin . After plating, the culture medium was changed every other day.
这个是别人论文中的方法。请问小鼠的操作哪些与大鼠不同？谢谢
