Engineering Adipose Tissue from Uncultured Human Adipose Stromal Vascular Fraction on Collagen Matrix and Gelatin Sponge Scaffolds

Abstract

Adipogenic potential was evaluated in uncultured stromal vascular fraction (SVF) loaded onto porous 3D collagen matrix and gelatin sponge scaffolds with predefined shapes. The SVF was isolated from 16 freshly lipectomized fat. Mean cell number was 6.0±4.68×10(7) cells/mL, and mean cell viability was 72%. Flow cytometric analysis revealed adipose-derived stromal cells (CD31(-), CD34(-/+), CD45(-), CD90(+), CD105(-), CD146(-), and CD166(+)) in the SVF. Three hours after harvest of fat, 200 μL of isolated SVF was loaded onto an experimental scaffold (4 mg in weight) and cultured in Dulbecco's modified Eagle's medium. Examination of the construct under an inverted light microscope and a scanning electron microscope demonstrated adequate seeding and active proliferation of the SVF cells on pore surfaces. Cells grew to varying sizes in clusters or in strands. On day 28, histologic study of the constructs by H&E staining revealed viable adipocytes in the microstructure. Positive Oil-Red O stain confirmed lipid accumulation in mature adipocytes. The presence of human adipocytes was further assessed by the presence of genomic DNA detected by GAPDH gene and by the mRNA expression of peroxisome proliferator activated receptor-γ, and lipoprotein lipase. The results demonstrated that new adipose tissue can be regenerated by seeding freshly isolated, uncultured SVF on 3D porous collagen matrix and gelatin sponge scaffolds.

Endothelial Differentiation of Human Stem Cells Seeded onto Electrospun Polyhydroxybutyrate/Polyhydroxybutyrate-Co-Hydroxyvalerate Fiber Mesh

“…Our results were in line with those of another study that seeded hASCs onto electrospun PLA nanofiber [48]. Furthermore, prior studies have demonstrated that hASCs can grow and proliferate with various types of biomaterials [49]–[52]. Thus, hASCs can be considered a good cell type for a tissue engineering approach, as they are derived from an abundant and available source and can be harvested with minimal morbidity.…”

Both Immediate and Delayed Intracavernous Injection of Autologous Adipose-derived Stromal Vascular Fraction Enhances Recovery of Erectile Function in a Rat Model of Cavernous Nerve Injury

“…Second, the exact composition of the rat SVF was not examined in this study. However, it is likely that the cell elements in the SVF of the rat are comparable to those in human SVF [30]. Third, we have not determined whether the decreased fibrosis in the corpus cavernosum is the result of a direct interaction between ADSC and the extracellular matrix or whether this is the result of increased innervation of the corpus cavernosum.…”

Secretory Factors From Rat Adipose Tissue Explants Promote Adipogenesis and Angiogenesis

“…For the in vivo study, gelatin sponge was used as scaffold material in adipose tissue engineering as reported elsewhere . Gelatin sponge (Nanjin Jinling Co., Nanjing, China) was punched into 2 × 2 × 0.25 cm 3 blocks under aseptic conditions.…”