Advances in Regenerative Medicine have shown a promising perspective in the use of mesenchymal stem cells in the treatment of various diseases and other conditions, such as in the cellular healing process. Mesenchymal stem cells are undifferentiated cells, with high proliferative capacity and potential differentiation in other cell lines, and when manipulated in vitro, can undergo several passages without losing their properties. The mesenchymal stem cells present some characteristics, such as adhesion to the plastic, the possibility of multiplying and specific surface markers expression pattern by flow cytometry labeling that allows its identification. However, since they are cultivated in vitro and are subjected to extensive manipulation, it is necessary to carry out various tests to prove their effectiveness and absence of abnormalities, such as chromossomic ones with cytogenetic analyses, which is essential to verify the safety and the chromosomal stability of mesenchymal stem cells. Therefore, the objective of the present study is to discuss the importance of cytogenetic through karyotype assay, in tissue engineering, in order to demonstrate cellular chromosomal stability for cultured samples of mesenchymal cells from dental pulp. The results showed chromosomal stability of the cells throughout the passages and after freezing.