Human adipose-derived stromal/stem cells are distinct from dermal fibroblasts as evaluated by biological characterization and RNA sequencing.
Mariane Izabella Abreu de MeloPricila da Silva CunhaMarcelo Coutinho de MirandaCamila Cristina Fraga FaracoJoana Lobato BarbosaAndrea da Fonseca FerreiraMarianna Kunrath LimaJerusa Araújo Quintão Arantes FariaMichele Ângela RodriguesAlfredo Miranda de GoesDawidson Assis GomesPublished in: Cell biochemistry and function (2021)
Human adipose-derived stromal/stem cells (ASC) have immunomodulatory properties and the potential to differentiate into several cell lines, important for application in regenerative medicine. However, the contamination with dermal fibroblasts (FIB) can impair the beneficial effects of ASC in cell therapy. It is then essential to develop new strategies that contribute to the distinction between these two cell types. In this study, we performed functional assays, high-throughput RNA sequencing (RNA-Seq) and quantitative PCR (qPCR) to find new markers that can distinguish ASC and FIB. We showed that ASC have adipogenic and osteogenic differentiation capacity and alkaline phosphatase activity, not observed in FIB. Gene expression variation analysis identified more than 2000 differentially expressed genes (DEG) between these two cell types. We validated 16 genes present in the list of DEG, including the alkaline phosphatase gene (ALPL). In conclusion, we showed that ASC and FIB have distinct biological properties as demonstrated by alkaline phosphatase activity and differentiation capacity, besides having different gene expression profiles. SIGNIFICANCE OF THE STUDY: Although many differences between stromal stem cells derived from human adipose tissue (ASC) and human dermal fibroblasts (FIB) are described, it is still difficult to find specific markers to differentiate them. This problem can interfere with the therapeutic use of ASC. This work aimed to find new markers to differentiate these two cell populations. Our findings suggest that these cells can be distinguished by biological and molecular characteristics, such as adipogenic and osteogenic differentiation, alkaline phosphatase activity and differential gene expression profiles. The DEG were related to the regulation of the cell cycle, development process, structural organization of the cell and synthesis of the extracellular matrix. This study helps to find new cellular markers to distinguish the two populations and to better understand the properties of these cells, which can improve cell therapy.
Keyphrases
- cell therapy
- stem cells
- single cell
- rna seq
- mesenchymal stem cells
- extracellular matrix
- endothelial cells
- high throughput
- bone marrow
- cell cycle
- gene expression
- nlrp inflammasome
- genome wide
- adipose tissue
- induced apoptosis
- induced pluripotent stem cells
- genome wide identification
- liver fibrosis
- pluripotent stem cells
- oxidative stress
- dna methylation
- risk assessment
- cell cycle arrest
- wound healing
- climate change
- insulin resistance
- high resolution
- type diabetes
- transcription factor
- high fat diet
- genetic diversity
- human health