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Thrombopoietin Combined with Early-Acting Growth Factors Effectively Expands Human Hematopoietic Progenitor Cells In Vitro

Institute of Transplantation Immunology and Medical Department A., The National Hospital, University of Oslo, Oslo, Norway

Key Words. Thrombopoietin • Hematopoietic progenitor cells • CD34 • Expansion • Stem cell transplantation • Megakaryocytopoiesis

Dr. Jan-Arne Hunnestad, Institute of Transplantation Immunology, The National Hospital, N-0027 Oslo, Norway.

Thrombopoietin (TPO) is established as a powerful stimulant of megakaryocyte differentiation and platelet production both in vivo and in vitro. In preparation for future transplantation of ex vivo expanded CD34⁺ hematopoietic progenitor cells (HPCs), we have examined the in vitro effect of TPO on cultures of HPC when combined with other early-acting hematopoietic growth factors (GFs) in an attempt to decrease post-transplant thrombocytopenia and accelerate engraftment. By adding TPO to all possible combinations of GM-CSF, IL-3, and c-kit ligand (CKL) in a suspension culture system, we found a significant increase in both relative and absolute numbers of cells in cultures containing TPO of the megakaryocytic lineage and CD34⁺ cells after 14 days of culture.

The most efficient GF combinations for expansion of cell populations of the megakaryocytic lineage and HPCs were TPO, GM-CSF, and CKL, which increased the number of cells of the megakaryocytic lineage 78 fold and the number of CD34⁺ cells 1.8 fold. The number of CD34⁺ cells decreased in the cultures containing GM-CSF and CKL with no TPO present, and the number of cells of the megakaryocytic lineage was increased merely 27 fold. Based on our findings, we suggest adding cells from HPCs expanded in cultures containing TPO, GM-CSF, and CKL to unexpanded stem cells for stem cell transplantation.

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