Bancel S, Hu W S
Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis 55455-0132, USA.
Biotechnol Prog. 1996 May-Jun;12(3):398-402. doi: 10.1021/bp960023a.
Macroporous microcarriers are often used to cultivate animal cells. The pores in the interior of the beads provide surface and space for cell growth. It is not clear how anchorage-dependent and suspension cells populated these microcarriers during cultivation. Confocal laser scanning microscopy was employed to perform time lapse observation of the cells in the interior. The structure of the bead was stained with fluorescein isothiocyanate for visualization, while the cells were stained with dialkyl indocarbocyanines for tracking over time. It was observed that mouse fibroblastic cells CRE BaG2 did not move extensively after initial attachment. Some cell divisions were observed during the course of the experiments, and essentially all cells remained viable throughout. Few hybridoma cells were deposited into the pores in the interior of the microcarriers. The results suggest that the occupancy of the internal volume by cells after prolonged cultivation is largely due to the growth of cells that are deposited in the interior as opposed to the migration of cells from the external surface into the interior. This method of observing cell behavior in a three-dimensional structure may find applications in other three-dimensional cell culture systems. The animation of time lapse sections is available on the worldwide web at http:/ www.cems.umn.edu/ approximately wshu_grp/acre/microcarrier.html.
大孔微载体常用于培养动物细胞。微珠内部的孔隙为细胞生长提供了表面和空间。目前尚不清楚在培养过程中贴壁依赖性细胞和悬浮细胞是如何在这些微载体上生长的。采用共聚焦激光扫描显微镜对微载体内部的细胞进行延时观察。用异硫氰酸荧光素对微珠结构进行染色以使其可视化，同时用二烷基吲哚羰花青对细胞进行染色以便随时间追踪。观察到小鼠成纤维细胞CRE BaG2在最初附着后没有大量移动。在实验过程中观察到一些细胞分裂，并且基本上所有细胞在整个过程中都保持存活。很少有杂交瘤细胞沉积到微载体内部的孔隙中。结果表明，长时间培养后细胞占据微载体内部空间主要是由于沉积在内部的细胞生长，而不是细胞从外表面迁移到内部。这种在三维结构中观察细胞行为的方法可能在其他三维细胞培养系统中得到应用。延时切片的动画可在全球网站http:/ www.cems.umn.edu/ approximately wshu_grp/acre/microcarrier.html上获取。
