Mapping the Fate of Progenitor Cells During in vitro Cardiogenesis and Neurogenesis

Injection of pluripotent stem-cell derived cardiomyocytes and neurons represents a promising strategy to treat tissue damaged in heart or neurodegenerative diseases and trauma. Current techniques have been able to generate human cardiomyocytes and neurons, but these populations are molecularly heterogenous, leading to variable functional properties and concerns over disruption of normal tissue function. The intermediate cells types that arise from differentiated stem cells in the process of creating cardiomyocytes and neurons are also heterogenous. Defining the progeny of these intermediate populations will improve methods for differentiating more homogenous cardiomyocyte and neuronal populations by allowing enrichment of intermediate cell types that give desired mature cell types. In order to study how to best generate homogenous populations for clinical treatment, we designed and carried out several potential experimental strategies. First, genetic lineage tracing methods were applied to investigate the cell fate decisions that control cardiogenesis and neurogenesis. Here, we are designing a system that combines lineage tracing with high dimensional single cell mass cytometry to provide an unprecedented level of detail into the heterogeneity seen during in vitro cardiogenesis and neurogenesis. Second, we are optimizing and investigating current neuronal differentiation protocols using mass cytometry to characterize the heterogeneity of final cellular populations, particularly after administration of known differentiation factors and comparing results with previously published literature.