Abstract 13784: Single Cell Profiling Enables Surface Marker Discovery in Cardiac Progenitor Cells
In tissues like the heart, a complex and heterogeneous organ, it is often difficult to purify and characterize different cell types. Many of the intracellular biomarkers for different populations are known, however, the distribution of extracellular markers for the same cell populations is not yet fully documented. In this study we used a high throughput single cell qPCR method on the embryonic hearts of transgenic mice with cardiac specific Nkx2.5 driven eGFP expression.
This technique permits robust gene profiling of cardiomyocyte progenitors for both know intracellular and potential extracellular biomarkers on single cell level. A mixed population of GFP positive cells was purified using FACS and single cells from embryonic day 9.5 and 11.5 were analysed for 48 genes. Following statistical analysis, candidate extracellular markers for different cell types were identified. Especially the integrin and connexin protein families showed promise in regard to correlation with different intracellular lineage markers. An early differentiated cardiomyocyte population, being Myl2 positive at E11.5, divided into a low and high expression profile. These populations correlate differently to the expression of Itga6, Itga1, Itga5 and Flk1. As this divergence of populations does not appear to be distinguishable at an earlier time-point, it could indicate the emergence of a more mature population. These findings were used to FACS candidate populations and characterize them using standard qPCR. The sorted cells expressed many mature markers that indicated the possibility of isolating and differentiating between committed atrial or ventricular myocytes. In conjunction with these techniques, immunofluorescence sections were made to identify the location of the different populations in vivo.
These results could potentially contribute to the identification of widely expressed, but unique, combinations of surface markers for different cardiomyocyte lineages. Additionally, the approach of single cell expression profiling has the capacity to expand our understanding of how cardiac cells differentiate and provides us with a tool for isolation and purification of pure populations that could be used in stem cell based therapies for heart disease.
- © 2013 by American Heart Association, Inc.