Development and Evaluation of Cell Culture Devices with the Gas-permeable Membrane

摘要

Single-use plastic culture devices are commonly used for two-dimensional (2-D) cell cultures. However, as oxygen is generally supplied by uni-directional diffusion from the top gas-liquid interface, 2-D cell cultures in the absence of medium flow may experience rapid exhaustion of dissolved oxygen at the bottom of culture wares with high cell densities. To overcome this issue, special cell culture devices utilizing the gas-permeable membrane at the bottom have been developed. The gas-permeable membrane can provide additional oxygen supply thorough rapid equilibration of dissolved oxygen at the bottom of culture ware, where cells are growing. In this study, using typical cell lines such as CHO and HEK-293A, cell growth, metabolic activity, and productivity were evaluated in gas-permeable culture devices compared to those from conventional gas-impermeable culture devices. The specific glucose consumption rate and the yield of lactate from glucose were decreased, revealing the effective utilization of glucose to produce ATP by enhanced supply of oxygen in gas-permeable culture devices. This effect was more clearly observed at in vivo-like pericellular conditions with low oxygen level (5% O-2 in the gas phase). The 2-D cell culture device with the gas-permeable membrane at the bottom has shown it's attractive performance for expansion of therapeutic cells that request pericellular conditions particularly, which is important for the maintenance of in vivo cellular activities, including differentiation and proliferation with appropriate cell growth, metabolic activity, and productivity as shown here.