Background- Experimental studies performed with human olfactory tissue-derived mesenchymal stem cells (OTD-MSCs) have shown that these cells can ameliorate nerve cell regeneration. However, three dimensional cell cultures with various supporting scaffolds are required in order to use OTD-MSCs in tissue engineering. Preparation of such scaffolds is both difficult and expensive. Developing a method of repairing nerve damage solely using stem cells without the need of any supporting material is important. Methods- First, human OTD-MSCs were isolated, cultured, and characterized. Next, consecutive passages were conducted to obtain multilayer cell growth. The resulting cell mass could be suitable for tissue engineering models as well as nerve cell or tissue regeneration studies in the future. Results- Viability and adhesive properties of the resulting cell mass were examined and found to be suitable for use in nerve tissue regeneration. Conclusion- It is suggested that an in vitro-produced OTD-MSCs mass can be applied to a very small damaged region and could have a high potential for microenvironment formation.