Abstract
Mesenchymal stem cell cultures from umbilical cord blood are obtained in conditions of oxygen tension and thiol concentration that are likely to cause a potent oxidative stress. The aim of this work was:
- to test the possibility of combining immunophenotyping with the determination of intracellular glutathione (GSH) on single cells.
- To check in cord blood cells the presence of gamma- glutamyltranspeptidase, an enzyme capable of making the effects of the catabolism of GSH pro-oxidant or anti-oxidant.
- To provide preliminary information on the influence of oxygen tension on the metabolism of low molecular weight thiols in culture.
The results showed that cord blood do contain GGT-positive cells, and
that this enzyme is present also in cells that appear in the earlier
phases of cell culture. Both GSH and cyteinylglycine (substrate and
product of the catalytic activity of GGT) are found in the supernatant
of umbilical blood cell cultures, and their concentration seems to be
influenced by modifying the oxygen concentration in the gaseous
phase of the culture environment. The single-cell determination of
GSH by CMFDA, combined with immunophenotyping by fluorescent antibodies revealed to be feasible, and might allow a detailed understanding of the redox-regulation mechanisms during differentiation of umbilical cord blood stem cells.
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