Abstract
Background: Transplantation of sheet-shaped autologous skeletal muscle-derived cells (SMDCs) has been shown to produce therapeutic effects via paracrine factors in ischemic cardiomyopathy. SMDCs are heterogeneous cell population consist of myoblasts and non-myogenic cells, and their detailed characteristic and paracrine factors has not been fully analyzed. We herein applied single-cell RNA sequencing (scRNA-seq) to profile the population and paracrine factors of SMDCs. Methods and Results: Primary SMDCs of human origin were subjected to scRNA-seq and divided into 5 populations, expressing myogenic markers differently. These included four myogenic populations with higher expression of MYF5 (myoblasts), myogenin (early differentiated cells), mitosis-related gene (proliferating cells), and MYL3 (moderately differentiated cells) and non-myogenic population with lower expression of myogenic genes. On the other hand, PAX7 , quiescent satellite cell marker, and MYH4 , terminally differentiated marker, were not expressed in any populations. Furthermore, regarding the expression of key angiogenic factors, VEGF and HGF were highly expressed in the non-myogenic population and HMGB1 was expressed in both populations. Moreover, analysis of the gene expression of angiogenic factors differentially expressed in each population indicated that myoblast population highly expressed PDGFA and CCN (Cellular communication network factor) 1 , and early differentiated population highly expressed CCN2 . Although these factors were expressed in other populations, their expression tended to be lower in the moderately differentiated cell population. Conclusion: Analysis of SMDCs using scRNA-seq revealed that they are heterogeneous cells consisting of multiple myogenic populations with different degrees of differentiation and cytokine production, suggesting that single-cell gene profiling may be useful for the evaluation on characters of effector cells.
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