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Establishment of protocol to enable on-site cryopreservation of fat for repeat procedures
Surucu Y, Chinnapaka S, Yang KS, Ejaz A, Rubin JP.
University of Pittsburgh, Department of Plastic Surgery, Adipose Tissue Stem Cell Center
2022-01-19
Presenter: Yusuf Surucu
Affidavit:
I certify that the material proposed for presentation in this abstract has not been published in any scientific journal or previously presented at a major meeting.
Director Name: J. Peter Rubin, MD, MBA
Author Category: Fellow Plastic Surgery
Presentation Category: Basic Science Research
Abstract Category: General Reconstruction
Introductions: Autologous fat transfer is an effective treatment for soft tissue reconstruction. The main challenge is that, on average, 63% of the graft volume takes, this necessitates repeat procedures. Therefore, preserving harvested tissue on-site for future injections is a clinical need. This study investigates different cryopreservation methods and applies the best results for a clinically usable device.
Methods: Different cryoprotectant combinations, freezing temperatures, and conditions were tested, and the outcome of the cryopreservation was assessed by measuring cell viability using trypan blue and Calcin-Am staining two days post freezing. In vitro validation of optimized conditions was tested for up to 3 months. For in-vivo testing, Nu/Nu athymic mice were used, and human fat cryopreserved for seven days, 21 days, three months, or 11 months was compared to fresh fat for graft weight and volume retention histology at nine weeks post graft. At +4oC three months, stored combination compared to fresh.
Results: A combination of 10% DMSO and 2% human serum albumin at -80oC provided optimum cryopreservation. We observed no significant differences in cell viability of cryopreserved fat for up to 3 months compared to the fresh fat. Cryopreserved fat grafts showed weight and volume retention and histological morphology comparable to fresh fat grafts. The mixture was storable.
Conclusion: The result of this study will enable the development of devices with clinically compatible appendages and a defined protocol for clinical use for long cryopreservation of fat tissue at -80oC within a closed system.
