Evaluation of Three Devices for the Isolation of the Stromal Vascular Fraction from Adipose Tissue and for ASC Culture: A Comparative Study

Abstract

Adipose-derived stem/stromal cells (ASCs) reside in the stromal vascular fraction (SVF) of adipose tissue (AT) and can be easily isolated. However, extraction of the SVF from lipoaspirate is a critical step in generating ASC, and semiautomated devices have been developed to enhance the efficacy and reproducibility of the outcomes and to decrease manipulation and contamination. In this study, we compared the reference method used in our lab for SVF isolation from lipoaspirate, with three medical devices: GID SVF-1™, Puregraft™, and Stem.pras®. Cell yield and their viability were evaluated as well as their phenotype with flow cytometry. Further on, we determined their proliferative potential using population doublings (PD), PD time (PDT), and clonogenicity assay (CFU-F). Finally, we checked their genetic stability using RT-qPCR for TERT mRNA assay and karyotyping as well as their multilineage potential including adipogenic, chondrogenic, and osteogenic differentiation. Our results demonstrate that all the devices allow the production of SVF cells with consistent yield and viability, in less time than the reference method. Expanded cells from the four methods showed no significant differences in terms of phenotype, proliferation capabilities, differentiation abilities, and genetic stability.

Development and Validation of a Fully GMP-Compliant Process for Manufacturing Stromal Vascular Fraction: A Cost-Effective Alternative to Automated Methods

“…In the few last years, a substantial number of automated and semi-automated devices have been introduced or are currently in development, with a total of 17 devices reported in the review of Oberbauer and colleagues [ 37 ]. A large body of articles have reported the biological characteristics of these devices [ 38 , 39 , 40 ], and in some cases, they use a manual method as reference [ 41 , 42 ]. However, the described manual protocols are not associated with any development data, leading to a high technical heterogeneity in the different steps of the processes and, consequently, in the biological results.…”

Uncultivated stromal vascular fraction is equivalent to adipose‐derived stem and stromal cells on porous polyurethrane scaffolds forming adipose tissue in vivo

“…As SVF cells yielded great adipogenic and angiogenic potential when applied without pretreatment, our data suggest that time‐consuming and cost‐intensive culture and induction is not necessary. Therefore, in view of the easier isolation protocols of SVF cells compared to ASCs and commercially available automated processing devices, this knowledge brings SVF‐based adipose tissue engineering closer to clinical applicability because a one‐step procedure can be achieved.…”

“…26 Underlying mechanisms include the natural interplay of endothelial cells, adipocytes, and stem cells and their secreted factors. 28 Therefore, SVF seems to be a promising cell source for translation into clinical practice, as its use is cost-effective, a one-step intraoperative application is possible, 29 devices for automatized procession exist, [30][31][32] and no adverse effects following patient application have been reported to date. 27 To our knowledge, no earlier investigation has compared ASCs and SVF cells with regard to their tissue formation capacities in adipose tissue engineering.…”

Current Applications of PRP and SVF Fractions in Stem Cell Therapy

“…The choice of adipose tissue harvest methods bothers on certain clinical parameters and as such a couple of options have been explored as seen in various papers that fulfilled (several criteria: Figure 1c) should be washed on the average of 3 times in 20mL syringes to remove lipoaspirate fluids, weighed and then digested in LR (1:1, v/v, fat/LR) containing collagenase NB6 at a final concentration of 0.1U/ml at 37 0 C for 45 min and under constant shaking (6).  Conventional method involving liposuction at -330mmHg into canister and no further manipulation in operation theatre prior to processing laboratory.…”

“…However, the quest for high quality, purified ASCs lead to development of semiautomated devices which been tested to augment efficiency and a reproducible end results. Currently the GID SVF-1™ has been demonstrated to recover more cells with preserved viability and cell phenotype using flow cytometry analysis (6).…”