Culture of Human Intestinal Cells in Simulated Microgravity

Abstract

We have used the rotating wall vessel (RWV) technique to culture standard cell lines as cell aggregates under conditions of very low fluid shear stress in aerobic and microaerophilic conditions (90% N2, 5% O2, and 5% CO2). The human intestinal adenocarcinoma cells lines Caco-2 and HT-29 in conventional two dimensional culture have been used for a wide range of research purposes. However, HT-29 cells when cultured in the RWV (Synthecon), formed large aggregates of up to 1 cm in their longest dimension. Haematoxylin and eosin stained sections of HT-29 aggregates showed large structures of cells with a small central zone of cell death compared to spheroids of HT-29 cells produced under standard two-dimensional conditions. Histochemical studies of the RWV aggregates (HT-29) showed populations of acid and basic mucin-secreting cells with basic mucin secretions predominant which encapsulated the aggregates. Caco-2 cells as expected did not show mucin secretion, but did show large vacuoles within RWV aggregates of about 30 cells which formed after 9 days in culture. Our work with the RWV system has demonstrated the value of this low shear stress system in enabling novel cell aggregate configurations with useful applications in the fields of pathogenesis of infection and tissue engineering.