Differential analysis of exosomes from adipose and umbilical cord-derived mesenchymal stem cell

Background: Mesenchymal stem cells have been extensively studied due to immunomodulation and tissue regeneration for therapeutic effects. The multipotent MSCs derived from bone marrow, adipose tissue, and umbilical cord have similar attractive properties and are widely studied in clinical trials. The advanced cell therapy in MSCs could also identify in the secretory compartment, especially in conditioned medium (MSC-CM) and exosomes derived from mesenchymal stem cells (MSC-Exo). Additionally, MSC cell-free products have an advantage for the skin regeneration effect without allogeneic cell transplantation.Objective: To analyze secretory compartments in the conditioned medium and MSC-exosome of the umbilical cord, and adipose tissueMethods: This was an experimental study of MSCs and its secretory compartment by isolation and Characterization of MSCs differentiation and flow cytometry: CD11b, CD34, CD45, CD73, CD79a, CD90, CD105, HLA-DR and collect condition medium to compare cytokine expression profiles especially focus anagen promoting factors, separation of exosome by ExoLutE kit and ultracentrifugation methods. Exosomes were characterized by nanoparticle tracking analysis and Flow Cytometry by CD9 andCD63. Proteomics was used to study exosomes.Result: Difference sources of exosomes showed varying sizes and numbers of particles. Using an ultracentrifuge and the ExoLutE kit, the size of exosomes produced by the ultracentrifuge method was smaller than the ExoLutE kit, and the concentration of exosomes extracted by the ultracentrifuge method appeared to be higher in exosomes derived from ADMSC than in exosomes derived from UCMSC. Important growth factors involving anagen factors were found Insulin-like growth factor 1 (IGF1), Vascular endothelial growth factor (VE GF), Platelet-Derived Growth Factor ( PDGF), Placental growth factor (PlGF), higher level in adipose mesenchymal stem cell-conditioned medium (ADMSC-CM) than umbilical mesenchymal stem cell-conditioned medium (UCMSC-CM) but Hepatocyte growth factor (HGF) was higher level in UCMSC-CM than ADMSC-CM. The Proteomics study showed that different pathways were adopted to explore the enrichment of proteins and this protein is not importantly associated with hair growth which has not been shown before effecting of hair growth from different techniques and sources of the exosome.Conclusion: This was the first study to analyze secretory compartments in the conditioned medium and MSC-exosome of the umbilical cord, and adipose tissue which isolation from different techniques by ExoLutE kit and ultracentrifugation methods. ADMSC-Exosome from ultracentrifugation has the largest number of exosome particles than other methods and no difference in concentration in UCMSC- Exosome between ultracentrifugation and ExoLutE kit. Optimistically, our findings were a good start toward revealing more certain essential growth factors and cytokines involving anagen factors for the hair cycle transported by MSC-derived exosomes from adipose tissue or umbilical cord which from the different techniques. Moreover, from the proteomics study, protein enriches in exosomes from this study are not importantly associated with hair growth which is caused by a low number of proteins to evaluate. There is also a need for more evaluation and investigation of the mechanism of each MSC-derived exosomes in vitro skin model for developing a new solution for the promising alternative cell-free-based therapeutic approach in Androgenetic alopecia.