Obtaining Human Umbilical Cord-Derived Mesenchymal Stem Cells and Cell Line Characterization: Immunophenotype, Multipotency and Cytogenetic Stability

Background Mesenchymal stem cells (MSCs) derived from the human umbilical cord offer a promising source for regenerative medicine due to their accessibility, low immunogenicity, and high differentiation potential. Unlike other types of MSCs, human umbilical cord MSCs (hUC-MSCs) are obtained through non-invasive procedures and raise fewer ethical concerns. This study aimed to represent our experience in isolating and characterizing hUC-MSCs in terms of their differentiation ability, immunophenotype, and genomic stability. Methods Human umbilical cord tissue was obtained with informed consent under aseptic conditions. MSCs were isolated using enzymatic disintegration of explants with collagenase I and cultured in Advanced DMEM/F12 supplemented with 2–10% fetal bovine serum. Cell morphology, proliferation rate, and viability were monitored during the first 10 passages of cultivation. Multipotency was assessed by inducing adipogenic, osteogenic, and chondrogenic differentiation using specialized culture kits, followed by histochemical staining with Oil Red O, Alizarin Red S, and Alcian Blue. Immunophenotyping was conducted via flow cytometry using antibodies against CD73, CD90, CD105 (positive markers), and CD34, CD45 (negative markers). Cytogenetic analysis was performed on passages 2, 4, 6, 8, 10 to assess genomic stability of cell line. Results Cultured hUC-MSCs displayed typical fibroblast-like morphology and adherence to plastic. Cells successfully underwent adipo, osteo, and chondrogeneic differentiation, confirmed by lineage-specific staining. Flow cytometry revealed a typical MSC immunophenotype (CD73+, CD90+, CD105+, CD34−, CD45−). Karyotype analysis demonstrated a normal diploid chromosomal number, confirming genomic stability during in vitro expansion, as well as stochastic occurrence of polyploid cells and cells with micronuclei at later passages (8–10), not indicative of genomic transformation. Conclusions Obtained hUC-MSCs exhibit stable genetic profiles, specific surface marker expression, and trilineage differentiation potential. These characteristics support their safety and effectiveness as candidates for future cell-based therapies and regenerative medicine applications.