Adipogenesis using human adipose tissue-derived stromal cells combined with a collagen/gelatin sponge sustaining release of basic fibroblast growth factor

作者：Ito Ran^*; Morimoto Naoki; Liem Pham Hieu; Nakamura Yoko; Kawai Katsuya; Taira Tsuguyoshi; Tsuji Wakako; Toi Masakazu; Suzuki Shigehiko

来源：Journal of Tissue Engineering and Regenerative Medicine, 2014, 8(12): 1000-1008.

DOI：10.1002/term.1611

• adipogenesis
• adipose tissue-derived stem cell (ASC)
• controlled release
• bFGF
• gelatin
• collagen
• scaffold

We have developed a collagen/gelatin sponge (CGS) that can provide a sustained release of basic fibroblast growth factor (bFGF). In our previous study, it was shown that CGS impregnated with the appropriate dosage of bFGF accelerates dermis-like tissue formation two or three times earlier than an existing collagen sponge. In this study, adipogenesis was evaluated using CGSs disseminated with adipose tissue-derived stem cells (ASCs). Human ASCs were primarily isolated from human adipose tissue that was obtained during breast cancer surgery with informed consent at Kyoto University Hospital. ASCs were isolated from collagenase digests of adipose tissue. ASCs were labelled with PKH26. CGSs (8mm diameterx3mm thickness) were impregnated with bFGF (0.1, 1, 7, 14 mu g/cm(2)) or normal saline solution. Then the labelled cells were disseminated (passage 3) on CGSs at a seeding density of 1x10(5) cells/cm(2) and implanted into the back subcutis of nude mice. Six weeks after implantation, adipogenesis at the administered site was evaluated. Immunohistological staining with von Willebrand factor (vWf) was performed to evaluate newly formed capillaries. Newly formed adipose tissue was observed macroscopically and histologically in all groups. The weight and area of regenerated adipose tissue were largest in the 1 mu g/cm(2) bFGF group. Under a fluorescent microscope, newly formed adipose tissue in the bFGF-administered group was PKH-positive. These findings show that ASCs differentiated and formed adipose tissue. In this study, we showed that our CGSs impregnated with bFGF could be used as scaffolds with ASCs for adipogenesis.

• 出版日期2014-12