Methodologic approaches for investigating human megakaryocyte development at a molecular level.

Abstract

Rapid advances in defining the extracellular regulators of megakaryocyte (MK) development have heightened interest in defining the biochemical and molecular mechanisms of action of these cytokines. The recent development of molecular micromethodologies such as in situ hybridization, the polymerase chain reaction, and the use of antisense oligodeoxynucleotides now make such studies possible in normal cells. To illustrate the utility of these methods, data gathered using these methods on developing normal human hematopoietic cells and MKs is presented. As might be expected, the results obtained demonstrate that growth factors have complex time and concentration effects on gene expression in morphologically recognizable human MKs. They also suggest that a more complete understanding of normal MK development at the molecular level will soon be possible.