Single Cell Analysis of the Developing Diaphragm at GD 13.5

We sequenced more than 13000 cells of the pleuroperitoneal folds of a mouse embryo to better understand the mosaic nature of the developing diaphragm. We identified 18 different clusters, and 10 unique cell types. Through our analysis, we were able to determine that most of the developing diaphragm is comprised of mesenchymal cells, and that genes necessary for proper diaphragm development are expressed in these types of cells. These observations provide a strong basis for furhter studies about the development and physiology of the diaphragm. We performed single cell RNA-seq on the developing diaphragm of a mouse embryo at GD 13.5, when the pleuroperitoneal folds are present. After meticulous dissection of the pleuroperitoneal folds from a mouse embryo, the tissue was dissociated into a single cell suspension. 13,482 cells were subsequently captured and used for cDNA library synthesis, sequencing, and cluster analysis. Expression information was recovered from each cell.