PMID: 11918560Mar 29, 2002Paper

Selective in vitro expansion and efficient retroviral transduction of human CD34+ CD38- haematopoietic stem cells

British Journal of Haematology
Yuk Yin NgFrank J T Staal

Abstract

Ex vivo expansion of primitive human haematopoietic stem cells (HSC) is clinically relevant for stem cell transplantation and gene therapy. Here, we demonstrate the selective expansion of CD34+CD38- cells from purified CD34+ cells upon stimulation with Flt3-ligand, stem cell factor and thrombopoietin. Over a 100-fold (range 80 to 128-fold) expansion of CD34+CD38- cells was observed with bone marrow and cord blood (CB). The expanded CD34+CD38- cells remained negative for lineage-specific markers and could be induced to differentiate into granulocytes, monocytes, megakaryocytes, erythrocytes, and T and B-lymphocytes in vitro. Lineage differentiation assays with single CD34+CD38- cells showed no loss of multilineage potential of expanded cells after ex vivo culture. We also demonstrated that the increase in frequency of CD34+CD38- cells was not as a result of the downregulation of CD38 expression during the culture. Quantitative analysis showed that the number of 6 week cobblestone area forming cells (CAFCwk6), a measure of proliferating HSC, in cytokine-stimulated CD34+ cells were increased by 20-fold. Expanded CD34+CD38- cells could be transduced efficiently with retroviruses encoding the low affinity nerve growth factor recepto...Continue Reading

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Citations

Apr 17, 2013·Cell Biology International·Xin YuHongwei Wang
Nov 27, 2004·Molecular Therapy : the Journal of the American Society of Gene Therapy·Francesca FicaraAlessandro Aiuti
Jul 16, 2005·Arteriosclerosis, Thrombosis, and Vascular Biology·Maarten B RookmaakerVictor W M van Hinsbergh

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