Abstract
BackgroundCryopreservation of fat grafts facilitates reinjection for later use. However, low temperature and thawing can disrupt tissues and cause lipid leakage, which raises safety concerns. Here, we compared the cryopreservation potential of stromal vascular fraction (SVF) gel processed from lipoaspirate with that of fat.MethodsHuman SVF gel and fat were cryopreserved at − 20 °C without cryoprotectant for 1 month. Fresh SVF gel and fat were used as controls. Tissue viability, adipose-derived stem cell (ASC) function, and the extracellular content were evaluated. At 3 months after transplanting the specimens to immunocompromised mice subcutaneously, the grafts were examined for retention, tissue engraftment, and inflammatory levels. The regenerative effect of cryopreserved SVF gel was evaluated in a murine ischemic wound healing model.ResultsAt 1 month, the cell death rate in the SVF gel group was 36 ± 2%. The survived ASCs not only could be isolated via explant culture but also preserved colony-forming and differentiation. However, prolonged cryopreservation exacerbated apoptosis. Assessment of recovered tissues showed that the morphology, cell viability, and extracellular protein enrichment were better in SVF gel-preserved tissues than in frozen fat. At 3 months after lipotransfer, the retention ability of 1-month cryopreserved fat was 41.1 ± 9% compared to that of 1-month cryopreserved SVF gel. Immunostaining results showed that adipose tissue regeneration and integrity in the 1-month cryopreserved SVF gel group were superior to those of the cryopreserved fat group. The cryopreserved SVF gel also accelerated healing of the ischemic wound, compared with cryopreserved fat.ConclusionCryopreserved SVF gel maintained tissue integrity and cell viability and resulted in a better long-term retention rate than that of cryopreserved fat. Cryopreserved SVF gel also showed superior regenerative potential and improved ischemic wound healing.
Highlights
Cryopreservation of fat grafts facilitates reinjection for later use
We hypothesized that removing adipocytes from fat tissue before cryopreservation may improve its tolerance during the cooling and recovery procedures, thereby increasing the viability of fat tissue and its regenerative potential
The extracellular matrix is preserved in stromal vascular fraction (SVF) gel after cryopreservation After cryopreservation for 1 month and thawing for 3 min, 0.1 mL of cryo-gel and cryo-fat were injected into a dish
Summary
Cryopreservation of fat grafts facilitates reinjection for later use. low temperature and thawing can disrupt tissues and cause lipid leakage, which raises safety concerns. Autologous fat grafting has significant therapeutic potential in revolumization and regenerative medicine [1]. The techniques for autologous fat transplantation have been optimized, they are associated with variable tissue resorption rates, requiring multiple surgeries to maintain the desired therapeutic effect [2, 3]. This limitation increased the scientific and clinical interest in the process of adipose tissue cryopreservation for reconstructive or regenerative medical purposes [4, 5]. Transplantation of cryopreserved fat tissue can lead to the formation of oil cysts. We hypothesized that removing adipocytes from fat tissue before cryopreservation may improve its tolerance during the cooling and recovery procedures, thereby increasing the viability of fat tissue and its regenerative potential
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