126 Adipose tissue engineering by human adipose-derived stromal cell seeded gelatin sponges

Abstract

The large bulk of lost craniofacial soft tissue is mainly adipose tissue. Current surgical approaches following tumor resection and trauma have limitations. Adipose derived-stromal cells (ADSCs), also named as preadipocytes, have been demonstrated to be a good cell candidate for adipose tissue engineering. However, characteristics of scaffold played important role in a successful tissue engineered adipose tissue. Objectives: The present study is to investigate whether in vitro adipogenesis could be induced both in a monolayer and 3-D by ADSCs. Methods: Healthy adult ADSCs were isolated by liposuction from fat pads. After discarding the unattached cells, the ADSCs were cultured in DMEM supplemented with 10% FBS and 1% antibiotics until they reached confluence. After trypsinization, ADSCs were subcultured at a density of 105 cells/100 mm-dish as a first passage. The monolayer and 3-D adipogenesis were assessed after seeding 105 ADSCs into each well of 6-well plates and 5 × 106 ADSCs/ml into biodegradable collagen sponges, respectively, followed by replacement of basic medium with adipogenic one. Results: In both monolayer and 3-D scaffolds, lipids containing adipocytes were observed 1 week after adipogenic stimulation, and further increased over the time. By contrast, no adipogenesis was observed in ADSCs cultured in basic medium. Conclusions: Human adipose-derived stromal cells isolated from fat pads can readily be induced toward adipogenesis in vitro. The in vivo adipogenesis potential of ADSCs is being investigated. (Supported by Whitaker Biomedical Engineering Foundation, March of Dimes Foundation)