Sustained expansion and transgene expression of coagulation factor VIII-transduced cord blood-derived endothelial progenitor cells.
OBJECTIVE
Although hemophilia A seems particularly suitable for gene therapy because even low amounts of plasma coagulation factor VIII (FVIII) provide a significant clinical benefit to the patients, the ideal target cell for recombinant FVIII expression and gene therapy approaches remains to be identified. In this study, we tested the capacity of cord blood-derived endothelial progenitor cells (CBECs) for FVIII expression on stable lentiviral transduction.


RESULTS
CD34+ endothelial progenitor cells (EPCs) from cord blood were differentiated into CBECs. Endothelial phenotype was characterized, and lentiviral transduction of early-passage CBECs with a vector encoding FVIII and EGFP did not alter their functional properties and proliferative potential. CBEC could be expanded by 5 to 9 orders of magnitude, thus allowing the expansion of up to 10(15) FVIII-secreting CBECs, starting from as little as 10(6) CD34+ cells. CBECs proved to be highly suitable for FVIII secretion, with 0.35 to 0.39 IU FVIII:C/5x10(4) cells per 48 hours (7.0 to 7.8 IU FVIII:C/10(6) cells per 48 hours), which remained stable over the expansion period.


CONCLUSIONS
Our data indicate that CBECs are attractive target cells for inherited coagulation disorders such as hemophilia A, which on lentiviral transduction can be readily expanded to large numbers of transplantable gene-modified cells in vitro.
Herder
Christian
C
Institute for Transfusion Medicine and Immunohematology, Red Cross Blood Donor Service Baden-Württemberg-Hessen, Sandhofstr. 1, 60528 Frankfurt am Main, Germany.
Tonn
Torsten
T
Oostendorp
Robert
R
Becker
Sven
S
Keller
Ulrich
U
Peschel
Christian
C
Grez
Manuel
M
Seifried
Erhard
E
D018952:Antigens, CD34 / Q000096:biosynthesis; D001777:Blood Coagulation / Q000235:genetics; D015153:Blotting, Western / Q000379:methods; D002454:Cell Differentiation / Q000235:genetics; D002455:Cell Division / Q000235:genetics; D002460:Cell Line; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry; D004730:Endothelium, Vascular / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D005169:Factor VIII / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D005312:Fetal Blood; D015316:Genetic Therapy / Q000379:methods; D049452:Green Fluorescent Proteins; D006412:Hematopoietic Stem Cells / Q000737:chemistry* / Q000378:metabolism* / Q000821:virology; D006467:Hemophilia A / Q000628:therapy; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics; D010641:Phenotype; D011993:Recombinant Fusion Proteins / Q000235:genetics; D014161:Transduction, Genetic / Q000592:standards / Q000706:statistics & numerical data; D019076:Transgenes / Q000235:genetics*
D018952:Antigens, CD34; D003470:Culture Media; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D005169:Factor VIII