The use of PCR to amplify the DNA or RNA of interest is highly advantageous since very few cells are required for analysis. Thus, by combining immunophenotyping with molecular biology, quantitative analysis of gene amplification and expression can be achieved in a specific cell population. Hematopoietic cells are derived from a small pool of primitive progenitor cells and differentiate from multipotential stem cells into committed progenitor cells that expand to functional end cells. Antibodies have been produced to some of the proteins on the membrane of hematopoietic cells. Since a unique repertoire of proteins is displayed for each subset along the differentiation pathway, we are able to separate and identify specific cell populations using Flow Cytometry and sorting.
Hematopoietic cells are responsive to a number of cytokines.
We hypothesize that hematopoietic cell subsets express a unique repertoire of cytokine receptors whose engagement by ligand regulates both proliferation and differentiation. This regulation can be directed by altering the combination of cytokines within the cell's immediate microenvironment. We have developed a process to simultaneously immunophenotype and molecular phenotype cells to evaluate cytokine receptor and ligand expression. Our main goal is to improve instrument performance. In the past, the emphasis of flow cytometry sorting has been on purity.