Phenotypical changes and proliferative potential of mesenchymal stem cells from humans Wharton’s jelly in the cultivation conditions

Abstract

Purpose. To develop a protocol for mesenchymal stem cells (MSCs) obtaining from wharton’s jelly of human umbilical cord, to study their phenotypic changes and proliferative potential in culture.Materials and methods. MSCs from human umbilical cord were cultured for 6 passages. To study morphological features of the cells in a culture we used adapted staining with eosin and gematoksilіn, Romanovsky–Giemsa, and toluidine blue. For nuclear DNA coloring we used intercalators: Hoechst 33342, and DARІ Ethidiumbromid. MSCs surface markers (CD105, CD90, CD73, CD34) expression was studied by FACS-analysis.Results. In the culture MSCs retain their morphological identity for 2 passages. Further a tendency to MSCs markers expression decreasing was observed, signs of culture degradation, spontaneous chondrogenic and adipogenic differentiation in late passages.Conclusion. Cultured MSCs of human umbilical cord for 2 passages retain their morphological characteristics and proliferative potential, further they lose multipotent mesenchymal phenotype.