Background Adipose tissue derived stroma cells (ASC's) offer for many advantages for tissue engineering strategies over mesenchymal stroma cells from other sources and ideal carrier materials have to be identified for them. The aim of this study was to demonstrate and to compare the effects of three clinically established biomaterials on proliferation and metabolic activity of rat ASC's in vitro. Materials and methods Rat adipose tissue derived stroma cells (ASC's) were isolated and differentiated into distinct lineages proved by lineage specific staining and gene expression analysis (RT-PCR). The biomaterials Bio-Gide®, Tutodent® Membrane and Belotero® Soft were tested with rat ASC's for their biocompatibility using scanning electron microscopy (SEM), cell vitality staining, cytotoxicity and proliferation tests (LDH, MTT, BrdU, WST-1). Results The collagen membrane Bio-Gide® resulted in a significantly higher viability and proliferation (WST-1, BrdU) compared to Tutodent ® Membrane. No significant difference was determined in the LDH and MTT test. The hyaluronic acid gel Belotero® Soft showed no cytotoxicity (LDH, FDA/PI) and had no negative effects on metabolic activity (WST-1, MTT) or cell proliferation (BrdU) of ASC's. Conclusion Our results indicate Bio-Gide® and Belotero® Soft as preferable carrier materials for ASC's. For the further establishment of ASC's-based treatment strategies, in vivo investigations on the tissue regeneration potential of these cell-biomaterial scaffolds should follow.