Single-cell RNA-Seq reveals the pseudo-temporal dynamic evolution characteristics of ADSC-induced differentiation into neurons

Abstract

Adipose-derived stromal cells (ADSC) has been frequently employed in the field of regenerative medicine. The molecular mechanism and genetic features of ADSC development into nerve cells, however, are unknown. This study used single-cell RNA sequencing(scRNA-seq) to reveal the features of gene expression changes during ADSC differentiation into neurons. We sequencd cells of ADSC group, the pri-1d group, and the induced 1h, 3h, 5h, 6h, and 8h groups using the BD Rhapsody platform. t-SNE ,Monocle2,GO,KEGG,and other algorithms were used to analyze sequence data. Results: From 7 groups, a total of 38453 cells were collected. 7 groups cells were divided into 0-13 clusters. ADSCs were located at the beginning of the trajectory by Monocle2 structured ,and the cells induced for 6h and 8h were largely dispersed in1st and 2nd branches of trajectory. The 5h-inducecells were primarily distributed in the trajectory' endpoints of 1st and 2nd branches. The GO items including cellular protein metabolism, cell adhesion, endocytosis, cell migration were enriched by up-regulated DEGs at 5h after induction. The KEGG analysis showed that induced 6h,8h groups mainly enriched pathways were oxidative phosphorylation, glutathione metabolism, Parkinson disease, Huntington disease, Alzheimer's disease, and other pathways. Conclusion: Two distinct cell state mechanisms primarily stimulate ADSCs to develop into mature neurons. By the fifth hour following induction, ADSCs had developed into mature neurons. The differentiated cells will experience aging-related degenerative changes if the induction response is kept up, and their physiological functions will also deteriorate.