Isolation of adipose-derived stem cells: a comparison among different methods

Abstract

Adipose-derived stromal cells (ASCs) are usually isolated by digestion with collagenase. We have compared alternative methods to isolate ASCs in a more economically viable protocol. Nine protocols using red blood cells lysis buffer solution, trypsin, collagenase and centrifugation were compared; the isolation rate, cell viability, expansion rate, immunophenotype and differentiation in adipogenic and osteogenic lineages were analyzed. ASCs were isolated and successfully maintained by digestion with trypsin. Cells presented similar immunophenotypes, adipogenic differentiation and in vitro proliferation but an osteogenic differentiation capacity up to seven times higher than ASCs isolated by collagenase. This alternative protocol is thus efficient and more cost-effective than the commonly-used methods and may represent a promising protocol for obtaining ASCs for bone tissue engineering.

A Standardized Method of Isolating Adipose-Derived Stem Cells for Clinical Applications

“…This method resulted in a fraction of stromal cells, including adipocytes, preadipocytes, endothelial cells, macrophages, fibroblasts, and a subpopulation of MSCs; although efficient, this procedure could be expensive and time-consuming when considered for clinical purposes. 10 With our method, neither animal-derived serum, medium of any type, red blood cell-buffered solutions or trypsin are necessary; moreover, the procedure is carried out step by step in a closed circuit, always preventing ASCs from air contact, thus minimizing any eventual risk of contamination.…”

“…This method resulted in a fraction of stromal cells, including adipocytes, preadipocytes, endothelial cells, macrophages, fibroblasts, and a subpopulation of MSCs; although efficient, this procedure could be expensive and time-consuming when considered for clinical purposes. 10 …”

Effect of autologous platelet-rich plasma on the chondrogenic differentiation of rabbit adipose-derived stem cells in vitro

“…Compared with BMSCs, ADSCs are superior in the range of sources, ease of obtaining and proliferative potential, and hold a broad future in cartilage regeneration (23,24). Currently, collagenase digestion is a commonly used method to obtain ADSCs, as ADSCs have very low density in adipose tissues which mainly include Col I (25). The isolated passage 0 ADSCs in the present study became adherent to the plastic plates within 24 h. The morphology of the passage 0 ADSCs was similar to that of fibroblasts with long spindle shapes.…”

The promising role for adipose-derived mesenchymal stem cells in tissue regeneration

“…Since lipoaspirate is readily collected during liposuction, tremendous numbers of ASCs can be relatively easily isolated from patients for use in therapeutic interventions [15,22]. Trypsin-mediated tissue digestion for ASC isolation may also provide an inexpensive method to harvest this cell population following lipoaspiration [23]. Successful culture of ASCs in xeno-free media without serum has also been achieved, further supporting consistent isolation and maintenance of ASCs in economically efficient in vitro conditions [24].…”