A protocol for primary isolation and culture of adipose-derived stem cells and their phenotypic profile
| Autor: | Adham F. Mohamed, Amira I. Fayad, Hadeer M. Rasheed, Myriam Helmy |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
business.industry
Mesenchymal stem cell Adipose tissue 030209 endocrinology & metabolism General Medicine Isolation (microbiology) Phenotype Cell biology 03 medical and health sciences 0302 clinical medicine Immunophenotyping Stem Cell Isolation Cell culture adipose-derived mesenchymal stem cells (ASCs) cell culture immunophenotyping mesenchymal stem cells (MSCs) Medicine 030212 general & internal medicine business |
| Zdroj: | Alexandria Journal of Medicine; Vol. 56 No. 1 (2020); 42-50 |
| ISSN: | 1110-0834 2090-2948 |
| Popis: | Background: Adipose tissue (AT) is a rich source of mesenchymal stem cells (MSCs), however, there is no standardized protocol for stem cell isolation and culture. This leads to inconsistency of the results and limits the comparison of the data from different laboratories. Our aim was to provide an applied protocol for ASCS isolation and expansion, study the cell behavior and define their cellular surface markers. ASCs were cultured from both resected adipose tissue (RAT) obtained following abdominoplasty or breast reduction and lipoaspirates (LPA) following laser-free liposuction. Method: the protocol entailed coculturing of stromal vascular fraction (SVF) with RAT as raw pieces using DMEM medium with varying glucose concentration. The coculture protocol aimed to mimic the normal physiological conditions required for cell growth. ASCs were immunophenotyped to define their MSCs surface markers by flowcytometry. Results: ASCs were isolated from coculturing RAT with SVF with fibroblast-like adherent cells morphology. The ASCs yield isolated from LPA was significantly greater than from RAT on day 14 and 28 (p = 0.002 |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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