title: Isolation of adipose tissue mesenchymal stem cells without tissue destruction: a non-enzymatic method.
abstract: The conventional enzymatic method is widely used for mesenchymal stem cells (MSCs) isolation from adipose tissue. The method holds major drawbacks; it is costly, time-consuming and results in a heterogeneous cell population. Besides, digestion of extracellular matrix causes cell injury and compromise proliferation and differentiation of the cells. Also, because of over handling the samples are also prone to contamination. Here, we introduce a non-enzymatic method for MSCs isolation without disturbing the cells habitat. Small pieces of adipose tissue obtained from animal or human liposuction were explanted into a culture flask, immobilized by fetal bovine serum (FBS) and incubated overnight. The explants were then irrigated with DMEM containing FBS. Within few days, the fibroblast-like cells migrated from the tissue and proliferated rapidly. When subconfluent, the cells were harvested, expanded through 3 passages and used for immunophenotyping and differentiation assays. As judged by flow cytometric analysis of surface markers (CD44(+), CD105(+), CD34(-), CD45(-)), Oil Red O and Alizarin Red staining, the MSCs isolated by our non-enzymatic method were pluripotent and exhibited the potential for differentiation into adipocyte and osteoblast. Great isolation yields, homogeneity of isolated cells, brief procedure, and high economy are the advantages of our method over the conventional protocol.
authors: Pharmacological Research Center of Medicinal Plants, School of Medicine, Mashhad University of Medical Sciences, Iran. Electronic address: ghorbania@mums.ac.ir.
authors: Ahmad
authors: A
authors: Ghorbani
authors: Immunogenetic and Cell Culture Department, Immunology Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
authors: Seyed Amir
authors: SA
authors: Jalali
authors: Department of Physiology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
authors: Masoumeh
authors: M
authors: Varedi
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology*; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D005434:Flow Cytometry / Q000379:methods; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D051381:Rats
keywords: Adipose; Liposuction; Stem cell; Tissue