OPTIMIZATION OF ISOLATION, LOW-SERUM CULTURING, AND IMMUNOPHENOTYPING OF HUMAN UMBILICAL CORD MESENCHYMAL STEM CELLS

Authors:

Abstract:

Mesenchymal stem cells from the human umbilical cord are a promising resource for cell therapy due to low immunogenicity and non-invasive harvesting. This study optimized an isolation protocol using collagenase I and evaluated cell properties during expansion. Umbilical cord explants were digested with 0.1 % collagenase I and cultured in Advanced DMEM/F12 with 2–10 % fetal bovine serum for 10 passages. Immunophenotypic analysis was performed via flow cytometry. Primary cells exhibited typical adherent fibroblast-like morphology. Low-serum maintenance (2 % fetal bovine serum) supported robust expansion without spontaneous differentiation. Flow cytometry confirmed high expression of positive markers (CD73, CD90, CD105) and complete absence of negative markers (CD34, CD45, HLA-DR). This protocol allows efficient isolation and expansion of human umbilical cord mesenchymal stem cells while maintaining their canonical immunophenotype under low-serum conditions, minimizing foreign protein exposure for future clinical applications.

Keywords:

mesenchymal stem cells umbilical cord cell culture immunophenotyping low-serum media flow cytometry

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Publication:

«World of Medicine and Biology» Vol. 22 No. 97 (2026), pp. 185-189
UDC 611.013.395:611.013.8

DOI:

https://doi.org/10.26724/2079-8334-2026-3-97-185-189