Tenogenic induction of human adipose-derived stem cells by soluble tendon extracellular matrix: composition and transcriptomic analyses

• DOI: 10.1186/s13287-022-03038-0
• Publication Date: 2022-07-29T13:18:12Z
• Authors: Ying Rao; Chenxian Zhu; Hoi Ching Suen; Shuting Huang; Jinyue Liao; Dai Fei Elmer Ker; Rocky S. Tuan; Dan Wang
• Keywords: Adipose-derived stem cells; Medicine (General); 0303 health sciences; Mass spectrometry; Tissue Engineering; Bioinformatics; Research; Stem Cells; RNA sequencing; Cell Differentiation; Extracellular matrix; QD415-436; Biochemistry; Extracellular Matrix; Tendons; 03 medical and health sciences; R5-920; Animals; Humans; Urea; Cattle; Collagen; Transcriptome; Tendon

Abstract Background Tendon healing is clinically challenging largely due to its inferior regenerative capacity. We have previously prepared a soluble, DNA-free, urea-extracted bovine tendon-derived extracellular matrix (tECM) that exhibits strong pro-tenogenic bioactivity on human adipose-derived stem cells (hASCs). In this study, we aimed elucidate the mechanism of tECM via characterization protein composition and comparison transcriptomic profiles hASC cultures treated with versus collagen type I (Col1) as control ECM component. Methods The was characterized by SDS-PAGE, hydroxyproline assay, proteomics analysis. To investigate action, differentiation tECM-treated compared serum medium or Col1-treated groups, assessed immunofluorescence for tenogenic markers RNA Sequencing (RNA-Seq). Results Urea-extracted yielded consistent composition, including collagens (20% w/w) at least 17 non-collagenous proteins (< 100 kDa) based MS Compared current literature, included key tendon components are functionally involved in regeneration, well those similar principal Gene Ontology (GO) functions (ECM-receptor interaction formation) signaling pathways focal adhesion). When used cell culture supplement, enhanced proliferation Col1 FBS treatment groups immunostaining tenogenesis-associated markers. Furthermore, RNA-Seq analysis revealed total 584 genes differentially expressed among three groups. Specifically, hASCs predominantly exhibited expression related ECM-associated processes, while mixture ECM- activity-associated which may explain part hASCs. Conclusions Our findings showed contained 20% w/w significantly enriched other components. treatment, supplementation induced distinct gene profiles. These provide insights into potential support development future tECM-based approaches repair.