Nerlich C, Langner A, Behrisch D, David H
Fachbereich Pharmazie, Bereiches Medizin (Charité), Humboldt-Universität zu Berlin.
Pharmazie. 1991 Jun;46(6):445-7.
In the present study rat hepatocytes isolated either by trypsin or by collagenase were investigated concerning its cultivability and its morphological properties. The cultivation was carried out as monolayer for 24 h. The yields of cells prepared by trypsin or collagenase amounted to 8 x 10(7) cells per liver and 15-30 x 10(7) cells per liver, respectively, with viabilities measured by trypan blue exclusion test of 70-80 and 90-95%, respectively. Nonhepatocytes were not taken into consideration. Using the electron microscopy it could be established that both freshly isolated and 24 h cultured hepatocytes were intact. There were no morphological differences between cells isolated by trypsin and cells isolated by collagenase. After 24 h cultivation hepatocytes prepared by trypsin showed a little tendency in forming a slightly flattened appearance and in forming intercellular contacts.
在本研究中,对用胰蛋白酶或胶原酶分离的大鼠肝细胞的可培养性及其形态学特性进行了研究。培养以单层形式进行24小时。用胰蛋白酶或胶原酶制备的细胞产量分别为每肝脏8×10⁷个细胞和每肝脏15 - 30×10⁷个细胞,用台盼蓝排斥试验测得的活力分别为70 - 80%和90 - 95%。未考虑非肝细胞。通过电子显微镜可以确定,新鲜分离的和培养24小时的肝细胞都是完整的。用胰蛋白酶分离的细胞和用胶原酶分离的细胞之间没有形态学差异。培养24小时后,用胰蛋白酶制备的肝细胞显示出有形成稍扁平外观和形成细胞间接触的轻微趋势。