| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1996: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Research Abstract |
To establish perfusion culture for efficiently colecting hematopoietic stem cells that are necessary for bone marrow transplantation, we investigated the optimal environment for growth of hematopoietic stem cells in vitro as well as the influence of HHV-6 on hematopoiesis as a representative mechanism of various clinically important viral infections.
During the past year, we demonstrated that HHV-6 directly inhibits the growth of erythroblasts or granulocyte and macrophage progenitors in a culture system using hematopoietic stem cells separated from cord blood. However, since thrombocytopenia is found in some children with exanthema subitum, an HHV-6 infection sometimes complicated by idiopathic thrombocytopenic purpura, a study on the growth of the megakaryocyte lineage is necessary. Delayd platelet recovery often becomes a clinical probelm in bone marrow transplantation, therefore, we attempted to culture megakaryocyte lineage hematopoietic stem cells (CFU-Meg), and to determine the optimal condition.
As the result of cultures under various conditions, when mononuclear cells from cord blood were cultured in medium containing 1% methylcellulose, 30% FBS,50 ng/ml TPO,2 mM 1-glutamine and 10^<-4> M 2-ME,formation of pellucid colonies characteristic of CFU-Meg occurred most efficiently after 10-12 days of culture. Though most of the colonies formed were CD41 positive and confirmed to be CFU-Meg, some colonies were CFU-M/GM,which are CD41 negative. Interaction between HHV-6 and other viruses and megakaryocyte progenitors is now under investigation.
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