Effective isolation of mesenchymal stem cells from human umbilical cord blood

Problem: Human umbilical cord blood (HUCB) has been poorly recognized as a rich source of Mesenchymal stem cells (MSCs). Background: HUCB still regarded as a waste postpartum tissue. However, HUCB contains Haematopoietic and non-Haematopoietic stem cells such as (MSCs). Haematopoietic stem cells (HSCs) have been isolated and used successfully in the clinical allogenic transplantation. Hypothesis: The aim of this study was to establish HUCB as a good source of MSCs using the novel clot spot method. Research: HUCB was collected (n=25) from candidates consenting women, who underwent elective caesarean section. HUCB clot was meticulously explanted in MesenCult basal medium and incubated in 5%CO2 incubator at 37OC. Qualitative and quantitative immunophenotyping of cells were achieved using fluorescein isothiocyanate (FITC) labelled antibodies reactive with CD34, CD45, CD29, CD44, CD73 and CD105 antigen markers. MSCs were examined in special media to be differentiated into neurogenic, osteogenic and adipogenic cells. Immunocytochemistry was used for embryogenic markers SOX2, Olig4, osteocalcin and FABP-4 identification Observations: MSC cultures using the clot spot method showed morphological changes in keeping with cell lineage differentiation. Within two weeks of culture, there is three -fold increase in cell number, compared with other methods. Cells were negative for HSC markers (CD34, D45), but strongly positive for MSC markers CD29 (97%), CD44 (95%), CD73 (95%) and CD105 (97%). MSCs expressed the neural SOX2, Olig4, osteogenic osteocalcin and adipogenic FABP-4 markers. Conclusions: Using novel techniques, it has been demonstrated that umbilical cord blood is a rich source for MSCs. Therefore, it should be considered as a good source of MSCs for potential use in clinical allogenic transplantatio