Marrow mesenchymal stem cells transduced with TPO/FL genes as support for ex vivo expansion of hematopoietic stem/progenitor cells.
A new marrow-derived mesenchymal stem cell (hMSC) line that could support expansion of hematopoietic stem/progenitor cells (HSPCs) was developed. Primary hMSCs were infected with retrovirus containing Flt-3 ligand and thrombopoietin genes. CD34+ cells from cord blood were expanded with primary hMSCs or transduced hMSCs. The expansion of total nucleated cells, CD34+ cells and mixed colonies containing erythroid and myeloid cells and megakaryocytes for 2 weeks coculture with transduced hMSCs was remarkably increased. The outputs of long-term culture-initiating cells for 2 and 4 weeks coculture with transduced hMSCs were also largely increased. The expansion rates of HSPCs with transduced hMSCs were unchanged for 6 weeks. In contrast, the expansion rates of HSPCs with primary hMSCs declined drastically through 6 weeks. SCID-repopulating cell expansion with transduced hMSCs for 4 weeks was significantly higher than that of uncultured CD34,+ cells and HSPCs expanded with primary hMSCs.
Xie
C-G
CG
Institute of Stem Cell Biology, College of Life Sciences, Zhejiang University, 232 Wen San Road, Hangzhou 310012, P. R. China.
Wang
J-F
JF
Xiang
Y
Y
Jia
B-B
BB
Qiu
L-Y
LY
Wang
L-J
LJ
Wang
G-Z
GZ
Huang
G-P
GP
D000328:Adult; D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation; D018920:Coculture Techniques; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics* / Q000502:physiology; D008648:Mesoderm / Q000166:cytology / Q000378:metabolism*; D008875:Middle Aged; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013926:Thrombopoietin / Q000235:genetics* / Q000502:physiology; D014161:Transduction, Genetic; D019076:Transgenes
D008565:Membrane Proteins; C084881:flt3 ligand protein; D013926:Thrombopoietin