Mechanical and Enzymatic Procedures to Isolate the Stromal Vascular Fraction From Adipose Tissue: Preliminary Results

Abstract

Adipose-derived MSCs (ASCs) and stromal vascular fraction (SVF) play an important role in regenerative medicine and in the treatment of osteoarthritis. ASCs extracted from lipoaspirates are a valuable cell source due to their abundance and accessibility. ASCs are retrieved from the aqueous fraction of the digested lipoaspirate. The aqueous fraction is known as SVF and includes, ASCs, endothelial precursor cells (EPCs), endothelial cells (ECs), macrophages, smooth muscle cells, lymphocytes, pericytes, as well as pre-adipocytes. To date, two types of techniques to isolate SVF have been proposed: enzymatic and mechanical. The enzymatic method is particularly indicated in SVF isolation since it disrupts the extracellular matrix (ECM) and the binding of adipocytes and other cells but is restricted by regulatory issues related to enzymatic procedures, especially within the European Community. Thus, making the search for alternative mechanical methods imperative. This study assesses the SVF harvested from subcutaneous abdominal fat via two different mechanical procedures and the standard enzymatic method to evaluate their eligibility in a clinical context. In particular, we analyze cell viability (at 0 and after 72 h) as well as the expression of cluster differentiation (CD) for each sample and the differentiation in adipocytic, chondrocytic, osteocytic linage. The mechanical procedures yielded no significant difference in cell viability and cluster differentiation pattern expression, even if enzymatic procedure still remain the “gold standard.” We retain that clinical efficacy in treating ostheoarthrosis with SVF administration is probably related to his anti-inflammatory and immunoregulatory effect, rather than the ability to differentiate in specific cell lineage. However, further studies are required to support and improve our findings.

Tissue Harvesting Site Effect on the Canine Adipose Stromal Vascular Fraction Quantity and Quality

“…Adipose tissue is an appealing cell source for regenerative and engineering medicine due to easy harvesting and the abundance of stem cell populations. Autologous adipose SVF injection has gained popularity in the orthopedic field because it is a favorable, minimally invasive, and non-surgical alternative for the handling of musculoskeletal disorders [ 28 ]. Despite the increasing number of pre-clinical and clinical studies on the potential role of SVF cells to treat osteoarthritis and/or cartilage lesions, clear findings are missing due to the insufficient standardization of SVF cell isolation and characterization [ 29 ].…”

Mesenchymal stromal cell products for intra-articular knee injections for conservative management of osteoarthritis

“…Adipose-derived stem cells (1–10% of the total nucleated cells) include hematopoietic stem cells, pericytes, supra-adventitial cells and mesenchymal stromal cells. 5 , 20 The characteristics of SVF, mainly cell availability, vary depending on whether the preparation protocol involves enzymatic digestion or mechanical breakdown of the lipoaspirate; 21 the former is termed cellular stromal vascular fraction of adipose tissue (cSVF) and the latter tissue stromal vascular fraction of adipose tissue (tSVF) or micro-fragmented adipose tissue. Although collagenase digestion provides a higher yield of nucleated cells, enzymatic manipulation represents a regulatory obstacle to protocol commercialization in most countries.…”

Adipocyte-Based Cell Therapy in Oncology: The Role of Cancer-Associated Adipocytes and Their Reinterpretation as Delivery Platforms

“…Regarding differences between cell types, a common strategy is to use conditioned culture media that will allow the growth of a single cell population. Rather than that, flow cytometry guarantees the distinction between cell types based on specific clusters of differentiation markers [149].…”