Claudia Cicione, Giuseppe Di Taranto, Marta Barba, Maria A. Isgrò, Alessio D’Alessio, Daniele Cervelli, Fabio V. Sciarretta, Sandro Pelo, Fabrizio Michetti, Wanda Lattanzi.
Plastic and Reconstructive Surgery, 2016 DOI: 10.1097/PRS.0000000000002014
The adipose stromal vascular fraction (SVF) has emerged as a rich and promising source of adipose derived stem cells (ASC).
ASCs exhibit high growth kinetics, plasticity and are able to induce efficient tissue regeneration in several biomedical applications without the need of culture expansion.
However, the required standards for clinical-grade cell manufacturing (eg., the current good manufacturing practices, cGMP, guidelines) prevent current experimental protocols for SVF isolation to be expanded into everyday clinical use.
This problem has been overcome with the development of a one-step procedure to treat different disorders through the use of
freshly harvested autologous SVF(Regenerative Fration). Here, we present an innovative device (myStem®), based on GMP-proof non-enzymatic tissue separation and cellular enrichment, enabling a rapid (10-15 minute) isolation of SVF from human lipoaspirates in a closed sterile system.
Adipose tissue (AT) from 5 donors was liposuctioned using the cannula contained in the kit and alternatively processed using: standard protocol (PLA), lipoaspirate fluid protocol (LAF) and MyStem® protocol.
All isolated cells were comparatively counted, analyzed morphologically, and characterized by flow cytometry. The harvested AT comprised small lobules, with intact cell membranes and structurally intact adipocytes. Histology showed the presence of cells without lipid content and placed in clusters along stromal axis of the lobule. The number of cells/gr isolated was respectively: 4×100.000 PLA, 1.8×100.000 LAF and 2×100.000 MyStem. Cells isolated with the three alternative protocols displayed a spindle shaped, bipolar morphology, comparable expression of surface antigens and differentiation capacity.
These results provided the first proof of principle on the feasibility of using a closed non-enzymatic protocol for collecting intact AT and separate a tissue fraction enriched in ASCs from a lipoaspirate sample. The isolated SVF display biological properties that can be easily exploited and translated in regenerative medicine applications.