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Cellular Optimization of Nanofat: Increased Cell Count and Viability Using LipocubeNano™

Introduction: Nanofat was introduced by Tonnard and Verpaele in 2013. Their initial observations in intradermal applications showed visual evidence of dermal regeneration. Since then, a number of Nanofat devices have been introduced. The cellular content in the processing of Nanofat is not the same in every device, yet the cellular composition forms the biologic action basis for Nanofat. We sought to find a different means to produce a matrix enriched Nanofat and optimize the cellular content.

Objectives: The primary objective of this study was to compare cell counts, cultures and cell viabilities produced by LipocubeNanoTM (Lipocube, Inc., London, UK) in comparison to Tulip’s Disposable NanoTransfer (Tulip Medical, San Diego, CA) processing methods, each having different tissue resizers and final filter sizes.

Methods: Twenty milliliters of fat was harvested from ten patients in order to test two methods of nanofat production. Ten milliliters of fat were used for each method and after the final product was obtained, an enzymatic digestion for SVF isolation was performed. A Muse Flow-cytometer was used to measure cell counts and cell viabilities. The characterization of ADSC (CD45-,CD90+/CD73+,CD90+), endothelial cells (CD45,CD31+), macrophages and monocytes (CD45+, CD14+) were performed by flow cytometry. The binding efficiency of the CD surface markers such as CD13, CD73, CD90, CD146 and CD34 were also examined by flow cytometry. Cells were then seeded in T-75 tissue culture plates (Proliferation medium; NutriStem® MSC XF Medium/serum free-Biological Industries) at 37°C, at 5% carbon dioxide. After 7 days, cell morphology was observed under light microscopy. We also compared theadipogenic differentiation capacity of ADSC in two groups. Gene expression profiles were examined by adipocyte-specific Adiponectin and Ppar genes.

Results: Cell number and cell viability results.

Cell Number/cc (n=10) Cell Viability (n=10)
Nanofat 1,44 x106 96,05%
LipocubeNano 2,24 x106 96,05%

Conclusion: Nanofat from LipocubeNanoTM has a higher regenerative cell count and more SVF than the other common mechanical method of Nanofat processing. This new means of mechanical processing, preserves more matrix, optimizing the cellular content of the Nanofat, thus having higher regenerative effect and differentiation potential.