Breaking down fat: Comparison of devices for SVF isolation

by Alexey Bersenev on January 14, 2015 · 0 comments

in adipose

Post to Twitter Send Gmail Post to LinkedIn

For the routine clinical use of adipose tissue-derived stromal vascular fraction (SVF), automated commercial processing devices should be considered. These devices allow to save a lot of time, money, decrease patient-to-patient and technician-to-technician variability, improve: safety of the procedure and quality of final SVF product. There is a variety of SVF devices worldwide in different stages of development and regulatory status. If you are trying to choose the best SVF processing device, you should consider the following paramenters:

  1. Device availability on a market
  2. Regulatory status (there are no SVF devices on US market, approved by FDA)
  3. Cost
  4. Functionality of the set and support (single use disposable, enzymes, centrifuge, other materials in the kit)
  5. Processing parameters: time, volume of input and output, automation, software customization
  6. Quality of SVF output

The cellular output of SVF processing device is the most important parameter. It includes: yield, viability, purity, cell composition, CFU (colony assay as surrogate for progenitor cell activity), residual enzyme level. There are only two available published studies, which compare different SVF processing devices. Let’s look at both of them.

The first study is by Aronowitz, published in mid of 2013. The authors compared 4 commercial SVF separation devices: (1) PNC’s Multi Station (manual device), (2) CHA Biotech Cha-Station, (3) Cytori Celution 800/CRS System, and (4) Medi-Khan’s Lipokit with MaxStem. Lipoaspirates from 5 patients were tested concurrently on 4 devices with following output assessment by “blinded” technicians. Use of Celution system resulted in better yield (number of nucleated cells per 1g of processed tissue). Viability: Celution 93%, Cha-Station 87%, Lipokit 72% and Multi Station 57%. Celution outperformed competitors in (1) CD34+%, (2) CFU assay and (3) residual enzyme level (lowest). Even though, Celution disposable kit was the most expensive ($1950, compare to $460-710 for other 3 devices), the authors argue that it could be economically more beneficial, based on yield of adipose stromal cells per dollar spent.

The second study – by Domenis, was published a week ago and freely available. The authors compared 3 commercially available devices: (1) Celution – Cytori, (2) Lipokit – Medikhan System and (3) Fastem – Corios (non-enzymatic method). The advantage of this study is a rigorous assessment of SVF output in vitro and clinically (as part of cell-assisted lipotransfer). Among in vitro tests – CFU, cell growth – expansion, single cell clonogenicity, multilineage differentiation, FACS and some others. Celution had better cell yield, Fastem was the worst. Celution and Lipokit had comparable % of CD34+ cells and CFUs. Fastem SVF had the highest % of CD45+ cells. Interestingly, after 1 weeks of culture, the authors detected expression of stem cell markers, such as Oct-4, Nanog and Sox-2 in all 3 tested outputs and non-enriched counterparts. With increasing culture time and passages, cells got similar “mesenchymal phenotype” (express CD44, CD73, CD90 and CD105). Unlike Fastem, Celution and Lipokit enriched cells were significantly increased in clonal ability (single cell colony assay), compare to non-enriched counterparts. Celution- and Lipokit-enriched cells had much better differentiation capacity than Fastem and non-enriched controls. Importantly, cell-assisted lipotransfer (using any of 3 processing methods) had better clinical results (in terms of graft stability/ thickness) than conventional lipotransfer in breast reconstitution patients. The most important message from the study – only enzymatic-based SVF isolation techniques, unlike mechanic methods, allow to enrich stem/ progenitor cells significantly.

Finally, I’d like to cite 2 more studies, which assessed devices in comparison to manual SVF isolation techniques. First study asseesed Incellator by Tissue Genesis: yield of nucleated cells 7.02×105 cells/ ml (similar to manual), viability 80.7% (similar to manual), enriched for CD34+ cells up to 70%. The second study validated Sepax2 by BioSafe: yield of nucleated cells 2.6×105 cells/ ml (62% > than manual process), viability >90% (same as manual), CFU 24% higher than in manual method.

If you had a chance to compare performance of 2 or more SVF devices, please share your experience!

***************
This post is a part of series Breaking Down Fat. In this series we will talk about identification, characterization and clinical processing of potentially therapeutic cell populations from adipose tissue. We started this series in response to the growing trend of wide (mostly uncontrolled) clinical use adipose-derived cells and some controversies/ misconceptions in the field.If you would like to contribute to this series or become a sponsor, please contact us!

{ 0 comments… add one now }

Leave a Comment

Previous post:

Next post: