Isolation and Characterization of Mesenchymal Stem Cells from Human Liposuction Aspirate

Abdelhamid, Alaa Eldin S; El Shahedy, Mohamed S; Mohammed, Mona H; Aly, Ghada S; Dessouki, Bassma M

doi:10.21608/scumj.2019.94981

Isolation and Characterization of Mesenchymal Stem Cells from Human Liposuction Aspirate

Document Type : Original Article

Authors

¹ Department of Clinical Pathology, Faculty of Medicine, Suez Canal University.

² Department of Virology, Faculty of Veterinarian Medicine, Suez Canal University.

³ Department of Anatomy and Embryology, Faculty of Medicine, Suez Canal University.

10.21608/scumj.2019.94981

Abstract

Background: Adipose derived stem cells (ADSCs) are a new type of mesenchymal stem cells (MSC) resemble bone marrow stem cells in their biologic activities. DScs is multipotent stem cells can differentiate into multiple cell types including osteoblasts, chondrocytes, adipocytes, myocytes, vascular endothelial cells, and neurons. Stromal vascular fraction (SVF) corresponds to ADSC and describes the cells obtained immediately from adipose tissues after the digestion of collagenase. Adipose tissue is a promising source of adult mesenchymal stem cells for therapeutic applications because it is available in large amounts (100 ml up to 1 liter) through liposuction and with minimal morbidity. Aim: To isolate and characterize the adipose-derived stem cells from human liposuction aspirate. Material and Methods: liposuction aspirate was used for the extraction of the stromal vascular fragment (SVF). SVF was used for culturing and isolation of hADSCs.ADSCs were cultured in tissue-culture flasks using collagenase digestion method. Flow cytometric analysis was used to identify ADSCs markers using fluorescence-activated cell sorting (FACS)against CD44, CD90, and CD45. Results: Human ADSCs were isolated successfully using collagenase digestion method. After 4 days the cells appeared to be spindle in shape and formed symmetric colonies. After 7 days, the cell sheet formed 60% confluence and completed to 90% confluence after 10 days incubation. FACS analysis of ADSCs surface markers revealed expression of CD44, and CD90 markers in most of the adherent cells. In contrast, most adherent cells were negative for CD45. Conclusion: Human ADSCs were isolated successfully from human lipoaspirate.
Keywords: ADSCs; MSC; SVF