Novel oxygenation system supports multilayer growth of HeLa cells.

We have developed a novel substratum in which gelatin is bonded to a reservoir of perfluorodecalin using a perfluoroalkylating technique. This forms a stable substratum supporting good adhesion for cells. HeLa cells cultured on this substratum continued to grow exponentially after the surface was covered with a monolayer forming a tissue-like structure of more than 19 layers of cells. Histological sectioning and staining of the block of tissue formed revealed the presence of mitotic figures deep within the structure. Every cell was surrounded by other cells, similar to growth of cells in vivo. This technique opens up a new approach to studying problems involved in cell-cell interaction and development of histotypic structures in vitro.