We compared migration, proliferation, growth curve, confluency and differentiation into adipogenic, osteogenic and chondrogenic cell lineages of mesenchymal stem cells derived from adipose tissue cultured in scratched and nonscratched cell culture flasks. Mesenchymal stem cells were isolated from rat adipose tissue using a nonenzymatic method. We investigated two groups. For the control group, minced adipose tissue was implanted conventionally onto the surface of standard plastic cell culture flasks. For the experimental group, the tissues were cultured in flasks with a scratched surface. We found that scratched flasks promoted cell migration, proliferation and confluency. Our findings suggest that scratched flasks may be used to ensure rapid, practical and safe isolation of adipose tissue-derived stem cells.