Investigate the stemness of adult adipose‐derived stromal cells based on single‐cell RNA‐sequencing

Abstract

The cellular heterogeneity and genetic features of stemness of adipose-derived stromal cells (ADSCs) remain unclear. Using single-cell RNA sequencing (scRNA-seq), we investigated the genomic features of the stemness gene in ADSCs with genetic variability. We cultured the ADSCs isolated from the fat waste of a healthy adult volunteers undergoing cosmetic plastic surgery to the third generation, used the BD Rhapsody platform to perform scRNA-seq, then used Monocle2 to analyze the growth and development trajectory of ADSCs, Cellular Trajectory Reconstruction Analysis Using Gene Counts and Expression (CytoTRACE) to evaluate the stemness gene characteristics in ADSCs clusters, and Beam to analyze the expression change characteristics of the main stemness related genes of ADSCs. According to the scRNA-seq data of 5325 ADSCs, they could be classified into nine cell clusters. According to CytoTRACE analysis, Cluster 3 of ADSCs had the highest stemness, whereas Cluster 8 had the lowest stemness.Pseudotime analysis revealed that Cluster 3 of ADSCs was primarily dispersed in the middle part of the growth and development trajectory, whereas Cluster 8 was primarily distributed at the end. We summarized the stemness of Cluster 3 in ADSCs with high expression of TPM1 and CCND1 genes in the metaphase of growth and development is the strongest, whereas the stemness of Cluster 8 with high expression of FICD, CREBRF, SDF2L1, HERPUD1, and HYOU1 genes in the telophase of growth and development is the weakest, providing a theoretical basis for screening and improving the therapeutic effect of ADSCs in cell transplantation research.

Genomic characteristics of adipose-derived stromal cells induced into neurons based on single-cell RNA sequencing

“…We kept cells with less than 10 % mitochondrial gene expression and more than 200 and less than 10,000 expressed genes. Mitochondrial genes were removed from the gene expression list [ [15] , [16] , [17] ].…”

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

“…The molecular process and genetic properties of ADSC development into nerve cells, however, are unknown. Previous research approaches and traditional transcriptome investigations have mostly been conducted at the population level of millions of cells, which cannot re ect cell heterogeneity, much alone explain the molecular change process of single cells [14][15][16]. With the rapid development of sequencing technology, cell capture, and bioinformatics, particularly the development of single cell RNA-sequencing (scRNA-seq), we now have a strong technical foundation to study cell heterogeneity and reveal the characteristics of gene expression changes in single cells under different states [17,18].…”