Innovative Adipose Tissue Fractionation for Transforming Fat into Specialized Components

Our research aims to fractionate adipose tissue to create targeted components and study their effect. Traditional fat graphing has issues like unpredictable absorption and uniform properties. Our protocol address this by creating specialized adipose components for targeted applications. Our lab will focus on how external factors, such as patient factors and surgical environment, affect adipose components transplantation.

[Narrator] To begin, allow the collected fat placed in a 60 milliliter syringe to stand undisturbed for 10 minutes to enable the liquid components to settle at the bottom. Carefully discard the liquid portion. Retain the collected fat in the 60 milliliter syringe, and centrifuge it at 1,200 G for three minutes at room temperature to further stratify the components, After centrifugation, discard the liquid portion at the bottom again and retain the upper fat layer now defined as Coleman fat. Now, transfer the upper two-thirds of the Coleman fat into a 60 milliliter syringe, defining it as low density fat. Then, transfer the remaining lower one-third into another 60 milliliter syringe, defining it as high density fat. For adipose matrix complex preparation, pass the high density fat through a filtering device with a sleeve and three internal sieves spaced at 1.5 millimeters. Collect the attached tissue and dehydrate it in a 100 mesh filter bag wrapped in gauze. Using forceps, determine dehydration is complete when no visible droplets fall from the tissue upon lifting. Define the final product as an adipose matrix complex. For stromal vascular fraction gel extraction, mechanically emulsify the low density fat by passing it between two syringes, connected by a female to female lure lock connector with a 1.4 millimeter internal diameter at a rate of 10 milliliters per second for one minute. Centrifuge the emulsified low density fat at 1,600 G for three minutes, and collect the middle gel-like layer as stromal vascular fraction gel. For adipose collagen fragment production, pulverize the low density fat using a specialized extractor at 30,000 revolutions per minute for 60 seconds to obtain a fat suspension. Transfer the fat suspension into a 20 milliliter syringe and attach a 0.25 millimeter unidirectional filter, push the piston, and collect the filtrate in the second syringe. Then, attach a 0.15 millimeter unidirectional filter to the second syringe, push the piston again, and collect the final filtrate in a third syringe. Finally, centrifuge the fat suspension at 3000 G for three minutes, and collect the solid portion at the bottom as adipose collagen fragment. The adipose matrix component, or AMC, exhibited significantly higher extracellular matrix and collagen content compared to Coleman fat, as shown by dense staining in histological and scanning electron microscopy images, and the collagen content in AMC was quantified at around 45% versus around 15% in Coleman fat mass. Masson staining showed that collagen was spread throughout the field in ACF, while in Coleman fat, it was limited to areas around fat cells. Bioinformatics and proteomic analysis of ACF identified a high abundance of plasma membrane and nuclear proteins and enrichment in collagen metabolism and organization processes with predominant presence of collagen types 1, 6, and 4.