Lodin Z, Hajková E, Hartman J, Faltin J, Srajer J
Physiol Bohemoslov. 1985;34(1):25-32.
Brain cells from 16 to 18-day-old mice embryos were dissociated by mild trypsinization and sieving. Immediately after dissociation the cells were preincubated in a PBS solution at -6 to +54 degrees C for 3 and 20 min. After this preincubation cells were rotated for 60 min at 37 degrees C in the PBS solution. Cellular adhesivity was estimated during this time period and EM pictures of organized in vitro aggregates after 24-28 h were taken. In a separate series of experiments, freshly dissociated were treated with DNAase before the rotation procedure. Preincubation in a cold or a warm medium did not alter the inhibition of cellular adhesivity significantly. Distinct inhibition of cellular adhesion was observed in cells preincubated above 53 degrees C. Adhesion was also inhibited below -5 degrees C, however, this effect was mainly dependent on the rate of freezing and thawing. Digestion of dissociated cells with DNAase (20 micrograms/ml) decreased cell adhesion. At 37 degrees C the adhesivity decreased by about 20%. Aggregates of cells preincubated at 0 degrees C for 20 min did not exhibit marked EM changes after 24-28 h in vitro. The present results have shown the rather high resistance of molecules responsible for cellular adhesion and its reversibility to temperature changes. Furthermore, non-specific cellular adhesion was shown on physically active DNA molecules.
通过温和的胰蛋白酶消化和筛分来分离16至18日龄小鼠胚胎的脑细胞。分离后，立即将细胞在-6至+54摄氏度的PBS溶液中预孵育3分钟和20分钟。预孵育后，细胞在37摄氏度的PBS溶液中旋转60分钟。在此期间评估细胞粘附性，并在24至28小时后拍摄体外有组织聚集体的电子显微镜照片。在另一系列实验中，新鲜分离的细胞在旋转程序之前用DNA酶处理。在冷或热培养基中预孵育不会显著改变细胞粘附性的抑制作用。在53摄氏度以上预孵育的细胞中观察到明显的细胞粘附抑制。在-5摄氏度以下粘附也受到抑制，然而，这种效应主要取决于冷冻和解冻的速度。用DNA酶（20微克/毫升）消化分离的细胞会降低细胞粘附性。在37摄氏度时，粘附性降低约20%。在0摄氏度预孵育20分钟的细胞聚集体在体外24至28小时后未表现出明显的电子显微镜变化。目前的结果表明，负责细胞粘附的分子具有相当高的抗性，并且其对温度变化具有可逆性。此外，在具有物理活性的DNA分子上显示出非特异性细胞粘附。
