Pluripotent Stem Cell Culture Scale-Out

Excerpt

Culture systems for human pluripotent stem cell (hPSC) expansion help enable the generation of a nearly unlimited pool of cells for use in downstream differentiation, disease modeling, drug discovery, and therapeutic applications. Monolayer culture is desirable in many circumstances due to the long history of validated assays and differentiation protocols. General culture protocols typically focus on methods for small plate formats (e.g., 6-well plate) yielding approximately 20 million hPSCs per plate. However, for some high-throughput screening and therapeutic applications, hPSCs expanded to a scale exceeding 1 billion cells may be required. This chapter provides a method for scale-out of hPSCs in monolayer cell culture conditions using commercially available cell culture media systems in conjunction with large surface area vessels. This scale-out procedure uses cell therapy grade reagents, which make application of cells and cell products to patients possible. It also provides insight into how to monitor cell health, assess pluripotency in normal stem cell cultures, and evaluates the capability to cryopreserve high cell densities.