An Optimized Method for Adipose Stromal Vascular Fraction Isolation and its Application in Fat Grafting

Author(s):  
Liang Cao ◽  
Feng Xiaoming ◽  
Qiang Zhang ◽  
Junbiao Fang ◽  
Chunhua Chu ◽  
...  
2018 ◽  
Vol 39 (6) ◽  
pp. NP213-NP224 ◽  
Author(s):  
Fang-Wei Li ◽  
Hai-Bin Wang ◽  
Jin-Ping Fang ◽  
Li Zeng ◽  
Chun-Lin Chen ◽  
...  

2011 ◽  
Vol 6 (2) ◽  
pp. 103-111 ◽  
Author(s):  
Valerio Cervelli ◽  
Pietro Gentile ◽  
Barbara De Angelis ◽  
Claudio Calabrese ◽  
Alessandro Di Stefani ◽  
...  

Author(s):  
Wenqing Jiang ◽  
Junrong Cai ◽  
Jingyan Guan ◽  
Yunjun Liao ◽  
Feng Lu ◽  
...  

Background: Autologous fat grafting has been a widely used technique; however, the role of adipose-derived stem cells (ASCs), extracellular matrix (ECM), and microenvironment in fat regeneration are not fully understood.Methods: Lipoaspirates were obtained and processed by inter-syringe shifting to remove adipocytes, yielding an adipocyte-free fat (Aff). Aff was then exposed to lethal dose of radiation to obtain decellularized fat (Df). To further remove microenvironment, Df was rinsed with phosphate-buffered saline (PBS) yielding rinsed decellularized fat (Rdf). Green fluorescent protein (GFP) lentivirus (LV-GFP)-transfected ASCs were added to Df to generate cell-recombinant decellularized fat (Crdf). Grafts were transplanted subcutaneously into nude mice and harvested over 3 months.Results: Removal of adipocytes (Aff) didn’t compromise the retention of fat grafts, while additional removal of stromal vascular fraction (SVF) cells (Df) and microenvironment (Rdf) resulted in poor retention by day 90 (Aff, 82 ± 7.1% vs. Df, 28 ± 6.3%; p < 0.05; vs. Rdf, 5 ± 1.2%; p < 0.05). Addition of ASCs to Df (Crdf) partially restored its regenerative potential. Aff and Crdf exhibited rapid angiogenesis and M2-polarized macrophages infiltration, in contrast to impaired angiogenesis and M1-polarized inflammatory pattern in Df. GFP + ASCs participated in angiogenesis and displayed a phenotype of endothelial cells in Crdf.Conclusion: Adipose ECM and microenvironment have the capacity to stimulate early adipogenesis while ECM alone cannot induce adipogenesis in vivo. By directly differentiating into endothelial cells and regulating macrophage polarization, ASCs coordinate early adipogenesis with angiogenesis and tissue remodeling, leading to better long-term retention and greater tissue integrity.


Author(s):  
Zhibin Yang ◽  
Shengyang Jin ◽  
Yu He ◽  
Xinyu Zhang ◽  
Xuefeng Han ◽  
...  

Abstract Background Over the past two decades, fat grafting has been extensively applied in the field of tissue regeneration. Objectives The authors investigated the therapeutic potential of microfat, nanofat and extracellular matrix/stromal vascular fraction gel (SVF-gel) in skin rejuvenation. Methods Microfat was harvested by a cannula with multiple 0.8 mm smooth side holes and processed with a fat stirrer to remove fibers. Nanofat and SVF-gel were prepared according to previously reported methods. We evaluated their structure and viability. Then, stromal vascular fraction (SVF) cells from the three types of samples were isolated and characterized, and the cell viability was compared. Results The microstructure of the three samples showed distinct differences. The microfat group showed a diameter of 100 to 120 .0μmunder the microscope and presented abotryoid shape under Calcein-AM/Propidium iodide (AM/PI) staining. Scanning electron microscopy (SEM) analysis showed that the microfat maintained integral histological structure.In the nanofat group, no viable adipocytes and no normal histological structure were observed, with high levels of free lipids.The SVF-gel group showed uniform dispersion of cells with different sizes and parts of the adipose histological structure. Cell count and culture revealed that the number of viable SVF cells decreased distinctly in the nanofat group compared with the microfat group. In contrast, the number of viable SVF cells in the SVF-gel group increased moderately. Clinical applications with microfat showed marked improvements in skin wrinkles. Conclusions The study showed that the microfat could preserve the integrity of the histological structure and presents the advantages of subcutaneous volumetric restoration and improvement of skin quality in skin rejuvenation compared with the nanofat and SVF-gel.


2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Jingwei Feng ◽  
Wansheng Hu ◽  
Mimi Lalrimawii Fanai ◽  
Shengqian Zhu ◽  
Jing Wang ◽  
...  

Abstract Background Cryopreservation 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. Methods Human 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. Results At 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. Conclusion Cryopreserved 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.


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