Combination use of dextran sulfate and polyvinyl alcohol prevent excess aggregation and promote growth of pluripotent stem cells in suspension culture

We report a new way for human pluripotent stem cells suspension culture. For clinical applications of cell-based therapies, a large quantity of human pluripotent stem cells (hPSCs) produced in standardized and scalable culture processes is required. Suspension culture for the increase in hPSCs has been one of the major challenges. Previously, Lipsit/Several groups reported that DS prevented aggregation of hPSCs and therefore regulated the aggregation size in a dose dependent manner. whereas, the yield of hPSCs is far from clinical needs. Here, by using cell aggregation assay, we developed methodologies for scaled up hPSCs production in stirred bioreactors. In this study, DS and PVA, affected hPSCs expansion and yield in stirred suspension cultures, were identified. Aggregates formed with these chemical compound kept the high expression rates of pluripotent marker genes and had the abilities of proliferation. These studies demonstrated that DS and PVA were useful for suspension culture for hPSCs and which remained pluripotent and capable of differentiating into all three germ layers. This knowledge will lead to the development of a simple and robust method for the mass culture of hPSCs.