Hartman J, Srajer J, Fleischmannová V, Lodin Z
Physiol Bohemoslov. 1981;30(4):295-305.
The adhesion of mouse embryonic brain cells was measured in a rotating chamber. A method is proposed for quantitative evaluation of adhesion kinetics. Dissociated cells were incubated in a planparallel chamber and pictures were taken between time 0-120 min. Film negatives were evaluated by computer--controlled scanning. Ten thousand individual data area obtained from one frame and 1,000 levels of absorbency are distinguished. A method is described which allows the discrimination of area, density and the shape of adhering cells. The influence of the dissociation procedure on cellular adhesion was studied. Short trypsinisation (0.025% trypsin for 5 min) followed by sieving was most favourable for adhesion. Mechanical sieving and dissociation with EGTA (Ca2+ chelator) gave less satisfactory results. Significantly diminished adhesion was observed after prolonged trypsinisation. If cells were incubated in media lacking Ca2+, adhesion was significantly inhibited. The kinetics of adhesion follows the curve of flocculation kinetics independently of the dissociation procedure and composition of the medium.
在旋转腔室中测量小鼠胚胎脑细胞的黏附情况。提出了一种定量评估黏附动力学的方法。将解离的细胞在平面平行腔室中孵育，并在0至120分钟之间拍照。通过计算机控制扫描对底片进行评估。从一帧中获取一万个单独的数据区域，并区分出1000个吸光度水平。描述了一种能够区分黏附细胞的面积、密度和形状的方法。研究了解离过程对细胞黏附的影响。短时间胰蛋白酶消化（0.025%胰蛋白酶处理5分钟）后进行筛选对黏附最为有利。机械筛选和用EGTA（钙离子螯合剂）解离的效果较差。长时间胰蛋白酶消化后观察到黏附显著降低。如果将细胞在缺乏钙离子的培养基中孵育，黏附则会受到显著抑制。黏附动力学遵循絮凝动力学曲线，与解离过程和培养基组成无关。
