• ISOLATION AND CHARACTERIZATION OF SECRETED EXOSOMES FROM ADIPOSE-DERIVED MESENCHYMAL STEM CELLS
  • Hoda Fazaeli,1 Azar Sheikholeslami,2 Naser Kalhor,3 Mohsen Sheikhhasan,4,*
    1. Department of Mesenchymal Stem Cells, Academic center for education, culture and research, Qom branch
    2. Department of Mesenchymal Stem Cells, Academic center for education, culture and research, Qom branch
    3. Department of Mesenchymal Stem Cells, Academic center for education, culture and research, Qom branch
    4. Department of Mesenchymal Stem Cells, Academic center for education, culture and research, Qom branch


  • Introduction: Mesenchymal stem cells are among the most common types of adult stem cells used in therapeutic medicine which have a high rate of differentiation and repair potential. Furthermore, these cells can secrete exosomes. One of the most important functions of these biological materials is the creation of cell-to-cell communication, which can be used as an appropriate mechanism to the gene or drug delivery in therapeutic medicine for diseases treatment. As a result, isolating and characterizing of exosomes could be important for achieving this goal. In this study, we will present to isolate and characterize the exosomes secreted from adipose-derived mesenchymal stem cells
  • Methods: After receiving the adipose sample from patients following liposuction and transferring it to the laboratory, mesenchymal stem cells were isolated from the adipose tissue using an enzymatic method. Briefly, after washing the adipose sample with PBS and removing the adipose tissue membranes and blood vessels, ADSCs were isolated using the collagenase digestion enzyme following 45–60 min incubation in a 37°C, 5 % CO2 cell culture incubator. Cells at the third passage were chosen, and CD90, CD44, CD34, and CD45 markers were characterized by flow cytometry. Then, conditioned medium obtained from adipose-derived mesenchymal stem cell at the third passage were used to exosome isolation. Exosome was isolated using exosome isolation kit according to the manufacturer's protocol. The size and morphology of exosomes isolated from adipose-derived mesenchymal stem cells were investigated using Transmission electron microscopy. Additionally, the concentration of exosomes was determined by the Bradford method.
  • Results: After culture and passage of the adipose-derived mesenchymal stem cells, conditioned medium obtained from adipose-derived mesenchymal stem cell at the third passage was collected following 48 hours of culture and exosomes were extracted. The electron microscope showed that the isolated exosomes had a spherical shape with a diameter of 40-150 nm. Also, the mean concentration of exosomes was 100 mg/ml.
  • Conclusion: Considering the important role of exosomes in therapeutic medicine, the method of isolation and characterization presented in this paper can be an effective and fundamental step for future research studies in the field of the gene or drug delivery for diseases treatment
  • Keywords: Exosome, Isolation, Characterization, Adipose-derived mesenchymal stem cell, Therapeutic medicine