Preparation of the Stirred-Tank Bioreactor for Large-Scale Production of Human Mesenchymal Stem Cells

To begin, perform two-point calibration on the pH probe with a bioreactor controller. Then, to assemble a bioreactor vessel, install parts such as stainless tubes and impellers onto the head plate according to the manufacturer's instructions. Attach a ready-to-use culture-disposable consumable kit to the appropriate stainless steel parts, tubes, and ports on the head plate following the manufacturer's guidance.

Upon assembling the vessel, perform an air-tightness check of the fully assembled vessel as specified by the manufacturer's instructions. Autoclave the vessel with the consumable kit at 15 PSI and 121 degrees Celsius for 60 minutes. After cooling the vessel to room temperature, insert the temperature probe into the thermowell tube and connect the cables for the monitor, pH probe, and DO probe from the controller to the respective parts on the vessel.

Wrap the heat mat around the vessel tightly. Log into the system of the bioreactor controller and enter the parameters into the system according to the manufacturer's instructions. Weld the outlet tube on the medium bottle to the inlet C-Flex tube on the bottle containing 10 grams of pre-sterilized W01 microcarriers for MSC.

Pump 500 milliliters of culture medium from the storage bag into the bottle of microcarriers W01.