Selective <i>in vitro</i> expansion and efficient retroviral transduction of human CD34<SUP>+</SUP> CD38<SUP>-</SUP> haematopoietic stem cells
BRITISH JOURNAL OF HAEMATOLOGY, cilt.117, sa.1, ss.226-237, 2002 (SCI-Expanded, Scopus)
- Yayın Türü: Makale / Tam Makale
- Cilt numarası: 117 Sayı: 1
- Basım Tarihi: 2002
- Doi Numarası: 10.1046/j.1365-2141.2002.03370.x
- Dergi Adı: BRITISH JOURNAL OF HAEMATOLOGY
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
- Sayfa Sayıları: ss.226-237
- Acıbadem Mehmet Ali Aydınlar Üniversitesi Adresli: Hayır
Özet
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 (CAFC(wk6) ), 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 receptor (LNGFR) marker gene (17% to 44%, mean 27%), resulting in long-lasting expression of retroviral-encoded genes in progeny HSC and differentiated progenitors. We conclude that the combination Flt3-ligand (FL), stem cell factor and thrombopoietin (TPO) induced strong ex vivo proliferation of CD34(+) CD38(-) cells and that the absolute number of expanded cells with stem cell activity increased substantially in this population.