A Novel Strategy for Simple and Robust Expansion of Human Pluripotent Stem Cells Using Botulinum Hemagglutinin

Clinical and industrial application of human pluripotent stem cells (hPSCs) has been hindered by the lack of robust strategies to sustain cultures in an undifferentiated state. Here, we describe a simple and robust method to culture and propagate hPSCs, which we anticipate will remove major roadblocks in investigating the basic properties of undifferentiated hPSCs and accelerate cell-based manufacturing. We also provide an overview of the use of botulinum hemagglutinin, an inhibitor of E-cadherin, to maintain and expand various hPSC lines in an undifferentiated state in different culture conditions. Hemagglutinin selectively removes cells that have lost the undifferentiated state, dissociates aggregates in situ, and is easy to use, scalable, and reproducible.