Identification of chemically-induced osteogenic cells (ciOG) through single cell RNA-seq

Here we report the reprogramming of human fibroblasts to produce chemically-induced osteogenic cells (ciOG), and explore the potential uses of ciOG in bone repair and disease treatment. A chemical cocktail of RepSox, forskolin, and phenamil was used for osteogenic induction of fibroblasts by activation of RUNX2 expression. Following a maturation, the cells differentiated toward an osteoblast phenotype that produced mineralized tissue. Single-cell RNA sequencing analysis of ciOG identified unique clusters of cells, among which the cluster of active cells related to osteogenic cells was ~49% of the cells, or ~80% of active cells; another cluster of active cells (~12% of the cells) was related to fibroblasts. The reprogramming trajectory over time revealed the transition from fibroblasts to mainly osteogenic cells or to an inactive state. ciOG identification through single cell RNA-seq