Edmondson J W, Bang N U
Am J Physiol. 1981 Jul;241(1):C3-8. doi: 10.1152/ajpcell.1981.241.1.C3.
Biochemical and morphological properties of rat hepatic parenchymal cells isolated without calcium were compared to cells isolated by adding calcium to the isolation medium at the time of addition of collagenase. Calcium contents of the two cell preparations were 4.5 +/- 0.3 and 10.5 +/- 0.5 nmol/mg dry wt, respectively (P les than 0.001). Magnesium content of both preparations was 37 nmol/mg dry wt. Potassium contents were 92 and 154 meq/l, respectively (P less than 0.001). Potassium content of calcium-deficient cells increased to 161 meq/l following incubation for 30 min in a medium containing 1.6 mM ionized calcium. When incubated in a medium containing a subphysiologic concentration of ionized calcium, calcium-deficient cells rapidly lost the ability to exclude trypan blue and to retain lactate dehydrogenase activity. As contrasted to calcium-sufficient hepatocytes, calcium-deficient cells failed to accumulate alpha-aminoisobutyric acid by active transport and lacked microvilli and nuclear contents. This study supports simultaneous addition of calcium and collagenase to the isolation medium as a means for preserving physical, functional, and morphological integrity of isolated hepatic parenchymal cells.
将无钙条件下分离的大鼠肝实质细胞的生化和形态学特性，与在添加胶原酶时向分离培养基中添加钙所分离的细胞进行比较。两种细胞制剂的钙含量分别为4.5±0.3和10.5±0.5 nmol/mg干重（P＜0.001）。两种制剂的镁含量均为37 nmol/mg干重。钾含量分别为92和154 meq/l（P＜0.001）。在含有1.6 mM离子钙的培养基中孵育30分钟后，缺钙细胞的钾含量增加到161 meq/l。当在含有亚生理浓度离子钙的培养基中孵育时，缺钙细胞迅速丧失排除台盼蓝和保留乳酸脱氢酶活性的能力。与钙充足的肝细胞相比，缺钙细胞不能通过主动转运积累α-氨基异丁酸，并且缺乏微绒毛和细胞核内容物。本研究支持在分离培养基中同时添加钙和胶原酶，作为保持分离的肝实质细胞的物理、功能和形态完整性的一种方法。
