Enriching hematopoietic, endothelial and mesenchymal functional progenitors by short-term culture of steady-state peripheral blood mononuclear cells obtained from healthy donors and ischemic patients.

BACKGROUND: Non-mobilized peripheral blood contains mostly committed cells with limited numbers of early progenitors.
OBJECTIVES: To enrich functional progenitor cells from healthy donors and ischemic heart disease patients by short-term culture of mononuclear cells with defined culture conditions.
METHODS: Mononuclear cells obtained from healthy donors and ischemic heart disease patients were cultured for7 days in a cytokine cocktail. We tested the multilineage differentiation capacities and phenotype of cultured cells.
RESULTS: The short-term culture (7 days) of all study groups with a defined cytokine cocktail resulted in two distinct cell populations (adherent and non-adherent) that differed in their differentiation capacities as well as their cell surface markers. Cultured adherent cells showed higher differentiation potential and expressed endothelial and mesenchymal fibroblast-like surface markers as compared to fresh non-cultured mononuclear cells. The non-adherent cell fraction demonstrated high numbers of colony-forming units, indicating a higher differentiation potential of hematopoietic lineage.
CONCLUSIONS: This study proved the feasibility of increasing limited numbers of multipotent progenitor cells obtained from the non-mobilized peripheral blood of healthy donors and ischemic patients. Moreover, we found that each of the two enriched subpopulations (adherent and non-adherent) has a different differentiation potential (mesenchymal, endothelial and hematopoietic).