Design of a Single Well Bioreactor-Incubator System with Sterile Nutrient Media Supply

Perfused bioreactors have been developed with different designs and with various cell lines such as fibroblasts, osteoblasts and C2C12 myoblasts. All these current systems are custom made and mostly capable for only one cell type. Small seeding spaces and chambers inhibit longer cell studies and the observation of cell interaction.
This study presents an advanced perfusion bioreactor which allows cell observation for longer than 5 days on an area of 2.8cm2 with a volume of 17ml. Initial experiments investigated homogeneity of the heat distribution, which is precisely controlled and stable over the length of the experiment. The addition of perfusion to the system results in the manual feeding process with modified F12 nutrient media being unnecessary. The perfusion and perfusion rate are user controllable up to 8ml/h. Different inlets allow cell seeding, cell feeding and chemical stimulation.
Since the metabolism by-products are diluted and removed by the flow of the perfused system, no inhibited growth occurs and the pH-value will maintain constant at 7.4 which removes the need for balancing the CO2 environment. Fully enclosed and sealed assemblies with a controlled hot plate can be used outside of the incubator and incorporated into the stage of a microscope to track and monitor cell growth.
The bioreactor chamber consists of three parts of transparent annealed cast Acrylic plus sealing material. Acrylic is chosen since it is machinable by laser cutting, which is a fast and easy method of manufacture. Due to the annealing process sterilizing by ethanol is possible. Heat distribution analysis was made with an IR-camera. And the pH was tested by indicator paper. The flow rate was set at 3.3ml/h. Computer simulations for flow and heat distribution and standard tests with cell cultures showed that a round bioreactor chamber design has advantages due to more uniform conditions.
To track the cells during their distribution and over their whole life cycle, a