Adeno-associated viral vector transduction of human mesenchymal stem cells

被引:74
作者
Stender, Stefan
Murphy, Mary
O'Brien, Tim
Stengaard, Carsten
Ulrich-Vinther, Michael
Soballe, Kjeld
Barry, Frank [1 ]
机构
[1] Natl Univ Ireland Univ Coll Galway, Regenerat Med Inst, Galway, Ireland
[2] Aarhus Univ Hosp, Dept Orthopaed, DK-8000 Aarhus, Denmark
关键词
mesenchymal stem cells; adeno-associated virus; transgene expression; multipotential activity;
D O I
10.22203/eCM.v013a10
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Mesenchymal stem cells ( MSCs) have received considerable attention in the emerging field of regenerative medicine. One aspect of MSC research focuses on genetically modifying the cells with the aim of enhancing their regenerative potential. Adeno-associated virus ( AAV) holds promise as a vector for human gene therapy, primarily due to its lack of pathogenicity and low risk of insertional mutagenesis. However, the existing data pertaining to AAV transduction of MSCs is limited. The objective of this work was to examine the efficiency and kinetics of in vitro transduction using AAV serotype 2 in human MSCs and to assess whether AAV transduction affects MSC multipotentiality. The results indicated that human MSCs could indeed be transiently transduced in vitro by the AAV2 vector with efficiencies of up to 65%. The percentage of GFP-positive cells peaked at 4 days post-transduction and declined rapidly towards 0% after day 8. The level of transgene expression in the GFP-positive population increased 4-fold over a 10,000 fold viral dose increase. This dose-response contrasted with the 200-fold increase observed in similarly transduced 293-cells, indicating a relatively restricted transgene expression in MSCs following AAV mediated gene delivery. Importantly, transduced MSCs retained multipotential activity comparable to untransduced controls.
引用
收藏
页码:93 / 99
页数:7
相关论文
共 34 条
[21]   Stem cell therapy in a caprine model of osteoarthritis [J].
Murphy, JM ;
Fink, DJ ;
Hunziker, EB ;
Barry, FP .
ARTHRITIS AND RHEUMATISM, 2003, 48 (12) :3464-3474
[22]   Reduced chondrogenic and adipogenic activity of mesenchymal stem cells from patients with advanced osteoarthritis [J].
Murphy, JM ;
Dixon, K ;
Beck, S ;
Fabian, D ;
Feldman, A ;
Barry, F .
ARTHRITIS AND RHEUMATISM, 2002, 46 (03) :704-713
[23]   Bone marrow cells regenerate infarcted myocardium [J].
Orlic, D ;
Kajstura, J ;
Chimenti, S ;
Jakoniuk, I ;
Anderson, SM ;
Li, BS ;
Pickel, J ;
McKay, R ;
Nadal-Ginard, B ;
Bodine, DM ;
Leri, A ;
Anversa, P .
NATURE, 2001, 410 (6829) :701-705
[24]   Multilineage potential of adult human mesenchymal stem cells [J].
Pittenger, MF ;
Mackay, AM ;
Beck, SC ;
Jaiswal, RK ;
Douglas, R ;
Mosca, JD ;
Moorman, MA ;
Simonetti, DW ;
Craig, S ;
Marshak, DR .
SCIENCE, 1999, 284 (5411) :143-147
[25]   Adeno-associated virus type 2-mediated transduction in primary human bone marrow-derived CD34(+) hematopoietic progenitor cells: Donor variation and correlation of transgene expression with cellular differentiation [J].
Ponnazhagan, S ;
Mukherjee, P ;
Wang, XS ;
Qing, KY ;
Kube, DM ;
Mah, C ;
Kurpad, C ;
Yoder, MC ;
Srour, EF ;
Srivastava, A .
JOURNAL OF VIROLOGY, 1997, 71 (11) :8262-8267
[26]   Potential of mesenchymal stem cells in gene therapy approaches for inherited and acquired diseases [J].
Reiser, J ;
Zhang, XY ;
Hemenway, CS ;
Mondal, D ;
Pradhan, L ;
La Russa, VF .
EXPERT OPINION ON BIOLOGICAL THERAPY, 2005, 5 (12) :1571-1584
[27]   Stress defense in murine embryonic stem cells is superior to that of various differentiated murine cells [J].
Saretzki, G ;
Armstrong, L ;
Leake, A ;
Lako, M ;
von Zglinicki, T .
STEM CELLS, 2004, 22 (06) :962-971
[28]   Marrow stromal cells for interleukin-2 delivery in cancer immunotherapy [J].
Stagg, J ;
Lejeune, L ;
Paquin, A ;
Galipeau, J .
HUMAN GENE THERAPY, 2004, 15 (06) :597-608
[29]   Membrane-associated heparan sulfate proteoglycan is a receptor for adeno-associated virus type 2 virions [J].
Summerford, C ;
Samulski, RJ .
JOURNAL OF VIROLOGY, 1998, 72 (02) :1438-1445
[30]   Characterization of multipotential mesenchymal progenitor cells derived from human trabecular bone [J].
Tuli, R ;
Tuli, S ;
Nandi, S ;
Wang, ML ;
Alexander, PG ;
Haleem-Smith, H ;
Hozack, WJ ;
Manner, PA ;
Danielson, KG ;
Tuan, RS .
STEM CELLS, 2003, 21 (06) :681-693