Genome wide gene expression profiling of Col2.3GFP+ enriched osteoblasts

We identified 593 candidate genes that are highly enriched in Col2.3GFP+ osteoblasts. Gene Ontology analysis revealed that these 593 genes are highly enriched in endochondral ossification, collagen fibril organization, positive regulation of osteoblast differentiation, and regulation of bone mineralization. The top 3 annotation terms are all associated with endoplasmic reticulum (ER) and Golgi vesicle transport. Furthermore, we have identified several vesicle trafficking genes with roles in osteoblast differentiation and function. Overall design: Using Col2.3GFP transgenic mice that express green fluorescent protein (GFP) in osteoblasts, we isolated Col2.3GFP+ enriched osteoblasts from three sources: 1) in vivo freshly isolated from mouse long bone; 2) ex vivo isolated from mouse bone chip culture; and 3) in vitro directed differentiation from mouse embryonic stem cells (ESCs). We performed RNA-sequencing and identified 593 overlapping genes that are highly enriched in Col2.3GFP+ osteoblasts from all three sources.