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Sustained expression of coagulation factor IX by modified cord blood‐derived mesenchymal stromal cells
Author(s) -
Dodd Megan,
MarquezCurtis Leah,
JanowskaWieczorek Anna,
Hortelano Gonzalo
Publication year - 2014
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.2769
Subject(s) - mesenchymal stem cell , cord blood , multiplicity of infection , adipogenesis , viral vector , stromal cell , ex vivo , microbiology and biotechnology , in vitro , biology , immunology , chemistry , andrology , cancer research , medicine , recombinant dna , gene , virus , biochemistry
Background Hemophilia B patients are subject to frequent and spontaneous bleeding caused by a deficiency of clotting factor IX (FIX). Mesenchymal stromal cells (MSCs) have been used in cellular therapies as a result of their immunomodulatory properties, the ability to home to sites of injury and their amenability to various ex vivo modifications, including lentiviral‐mediated gene transfer. Methods MSCs were isolated from human umbilical cord blood and differentiated into adipogenic, chondrogenic and osteogenic lineages. A lentiviral DNA vector containing the human FIX gene was generated using traditional restriction enzyme digest and ligation techniques to generate viable replication‐incompetent lentiviral particles that were used to transduce MSCs. Quantitative measurement of FIX expression was conducted using an enzyme‐linked immunosorbent assay. Results The over‐expression of FIX was sustained in vitro at levels > 4 µg/10 6 cells/24 h and FIX coagulant activity was > 2.5 mIU/10 6 cells/24 h for the 6‐week duration of study. Lentiviral modification of cells with a multiplicity of infection of 10 did not adversely affect the potential of cord blood (CB) MSCs to differentiate to adipocytes, chondrocytes and osteoblastic cells, and the expression of functional FIX was sustained after differentiation and was similar to that in nondifferentiated cells. Conclusions Modification of human CB MSCs with a lentiviral vector resulted in sustained high FIX expression in vitro after differentiation to adipogenic, chondrogenic and osteoblastic cells. These modified MSCs could have applications in cellular therapies for hemophilia B. Copyright © 2014 John Wiley & Sons, Ltd.