Forsby N, Collins V P, Westermark B
Acta Pathol Microbiol Immunol Scand A. 1985 Sep;93(5):235-49. doi: 10.1111/j.1699-0463.1985.tb03947.x.
Using three lines of human normal glial cells and four established lines of human malignant glioma cells we have studied cell spreading following seeding onto glass and plastic substrata. The cells were detached with EDTA and trypsin, suspended in EMEM with 10% calf serum and studied with time-lapse, phase-contrast cinematography in suspension and during attachment and spreading. Cells were fixed and prepared for light microscopy while in suspension and during the spreading process. They were also prepared for scanning and transmission electron microscopy at different times during spreading. The projected areas of stained cells, in suspension and at different stages of spreading, were measured morphometrically and the results compared statistically. The glial cells in suspension were often found to retain somewhat their shape from the previous monolayer. They spread radially outwards with even lamellar cytoplasm and peripheral ruffling, as a group more quickly than the malignant glioma cells. They also became polarized and started to translocate in a shorter time. The glioma cells were spherical in suspension and characterized by pronounced blebbing of the cell surface. Blebbing continued during spreading and was finally replaced by ruffling at the edge. The cells spread like the glial cells radially outwards but the lamellar cytoplasm was occasionally somewhat irregular. Cells from the glioma lines spread as groups slower than the glial cells but with individual rates for the different lines. One of the glioma lines appeared to spread more thinly than the glial cells. Cells which sedimented on top of other cells could not spread. Aggregations of cells spread and became polarized more quickly than single cells in all cases.
我们使用三株人正常神经胶质细胞系和四株已建立的人恶性神经胶质瘤细胞系，研究了接种到玻璃和塑料基质上后的细胞铺展情况。用乙二胺四乙酸（EDTA）和胰蛋白酶将细胞消化下来，悬浮于含10%小牛血清的伊格尔最低必需培养基（EMEM）中，并通过延时相差显微镜摄影技术对其悬浮状态以及贴壁和铺展过程进行研究。在悬浮状态和铺展过程中，将细胞固定并制备用于光学显微镜观察。在铺展的不同时间点，还对细胞进行处理以用于扫描电子显微镜和透射电子显微镜观察。通过形态计量学方法测量悬浮状态下以及铺展不同阶段染色细胞的投影面积，并对结果进行统计学比较。悬浮状态下的神经胶质细胞通常仍保留其先前单层状态时的一定形状。它们呈放射状向外铺展，细胞质呈均匀的片状，周边有褶皱，作为一个群体，其铺展速度比恶性神经胶质瘤细胞更快。它们也会发生极化，并在更短的时间内开始移位。神经胶质瘤细胞在悬浮状态下呈球形，其特征是细胞表面有明显的泡状凸起。在铺展过程中泡状凸起持续存在，最终边缘处被褶皱所取代。这些细胞像神经胶质细胞一样呈放射状向外铺展，但片状细胞质偶尔会有些不规则。神经胶质瘤细胞系的细胞作为一个群体铺展速度比神经胶质细胞慢，但不同细胞系有各自的铺展速率。其中一个神经胶质瘤细胞系的铺展似乎比神经胶质细胞更稀疏。沉积在其他细胞之上的细胞无法铺展。在所有情况下，细胞聚集体比单个细胞铺展并极化得更快。
