Hierarchical structure of human megakaryocyte progenitor cells.

Abstract

Megakaryocytopoiesis is a complex biological process involving a series of cellular events that begins with the pluripotent hematopoietic stem cell and ultimately results in the biogenesis of platelets. A hierarchy of megakaryocyte (MK) progenitor cells has been previously defined based upon studies of in vitro megakaryocytopoiesis. Ontogeny-related changes in MK progenitor cells were analyzed in order to further define this cellular hierarchy. Unifocal colony-forming unit-megakaryocyte (CFU-MK)-derived colonies cloned from fetal bone marrow (FBM) formed after fewer days of in vitro culture and were 2.6-fold larger than those colonies cloned from adult bone marrow (ABM). The frequency of CFU-MK-derived colonies cloned from ABM was significantly greater. MK colonies, however, cloned from FBM morphologically consisted of both pure MK colonies and mixed colonies containing MKs, in which a core of CD41- cells were surrounded by CD41+ MKs. Large colonies resembling the primitive BFU-MK also were assayed from both FBM and ABM. These BFU-MK-derived colonies appeared after fewer days of incubation when FBM was assayed, compared to ABM, but at a significantly lower frequency. In addition, large unifocal MK colonies consisting of >300 cells (300-1000) appeared from cells cloned from fetal, but not adult, marrow. This type of colony represents a unique type of MK progenitor cell, termed the high-proliferative-potential cell-MK. Such colonies represent the progeny of the most primitive human MK progenitor cell identified to date. We also attempted to investigate the process of commitment of stem cells to the MK lineage. We explored the actions of thrombopoietin (TPO) on primitive hematopoietic cells in order to gain an understanding of stem cell commitment. CD34+ Thy-1+ Lin- marrow cells, which are enriched for pluripotent hematopoietic stem cells, were shown to express c-Mpl by the polymerase chain reaction. In addition, TPO alone was capable of inducing CD34+ Thy-1+ Lin- cells after two to three weeks to produce progeny composed entirely of MKs. These studies indicate that TPO has a profound effect on hematopoietic stem cells, and that the hierarchy of MK progenitor cells begins with the pluripotent hematopoietic stem cell.