Early osteoblastic differentiation induced by dexamethasone enhances adenoviral gene delivery to marrow stromal cells

Journal of Orthopaedic Research - Tập 22 - Trang 411-416 - 2004
Jeremy S Blum1, M Brandon Parrott2, Antonios G Mikos3, Michael A Barry1,2,4,5
1Department of Bioengineering, Rice University, Houston, TX, USA
2Department of Immunology, Baylor College of Medicine, Houston, TX, USA
3Department of Bioengineering Rice University Houston TX USA
4Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, TX, USA
5Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA

Tóm tắt

AbstractWe investigated the implications of induced osteogenic differentiation on gene delivery in multipotent rat marrow stromal cells (MSCs). Prior to genetic manipulation cells were cultured with or without osteogenic supplements (5 ± 10−8 M dexamethasone, 160 μM l‐ascorbic acid 2‐phosphate, and 10 mM β‐glycerophosphate). Comparison of liposome, retroviral, and adenoviral vectors demonstrated that all three vectors could mediate gene delivery to primary rat MSCs. When these vectors were applied in the absence or presence of osteogenic supplements, we found that MSCs differentiated prior to transduction with adenovirus type 5 vectors produced a 300% increase in transgene expression compared to MSCs that were not exposed to osteogenic supplements. This differentiation effect appeared specific to adenoviral mediated gene delivery, since there was minimal increase in retroviral gene delivery and no increase in liposome gene delivery when MSCs were treated with osteogenic supplements. In addition, we also determined this increase in transgene production to occur at a higher concentration of dexamethasone (5 ± 10−8 M) in the culture medium of MSCs prior to adenoviral transduction. We found that this increased transgene production could be extended to the osteogenic protein, human bone morphogenetic protein 2 (hBMP‐2). When delivered by an adenoviral vector, hBMP‐2 transgene production could be increased from 1.4 ng/105 cells/3 days to 4.3 ng/105 cells/3 days by culture of MSCs with osteogenic supplements prior to transduction. These results indicate that the utility of MSCs as a therapeutic protein delivery mechanism through genetic manipulation can be enhanced by pre‐culture of these cells with dexamethasone. © 2003 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved.

Tài liệu tham khảo

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Journal of Orthopaedic Research

Tập 22

411-416

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