Sustained and promoter dependent bone morphogenetic protein expression by rat mesenchymal stem cells after BMP-2 transgene electrotransfer

Abstract

Document VersionPublisher's PDF, also known as Version of Record (includes final page, issue and volume numbers) Please check the document version of this publication:• A submitted manuscript is the author's version of the article upon submission and before peer-review. There can be important differences between the submitted version and the official published version of record. People interested in the research are advised to contact the author for the final version of the publication, or visit the DOI to the publisher's website.• The final author version and the galley proof are versions of the publication after peer review.• The final published version features the final layout of the paper including the volume, issue and page numbers. Link to publication Citation for published version (APA):Ferreira, E., Potier, E., Vaudin, P., Oudina, K., Bensidhoum, M., Logeart-Avramoglou, D., ... Petite, H. (2012). Sustained and promoter dependent bone morphogenetic protein expression by rat mesenchymal stem cells after bmp-2 transgene electrotransfer. European Cells and Materials, 24, 18-28. General rightsCopyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.• Users may download and print one copy of any publication from the public portal for the purpose of private study or research.• You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal ? Take down policyIf you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. AbstractTransplantation of mesenchymal stem cells (MSCs) with electrotransferred bone morphogenetic protein-2 (BMP-2) transgene is an attractive therapeutic modality for the treatment of large bone defects: it provides both stem cells with the ability to form bone and an effective bone inducer while avoiding viral gene transfer. The objective of the present study was to determine the influence of the promoter driving the human BMP-2 gene on the level and duration of BMP-2 expression after transgene electrotransfer into rat MSCs. Cytomegalovirus, elongation factor-1α, glyceraldehyde 3-phosphate dehydrogenase, and beta-actin promoters resulted in a BMP-2 secretion rate increase of 11-, 78-, 66-and 36-fold over respective controls, respectively. In contrast, the osteocalcin promoter had predictable weak activity in undifferentiated MSCs but induced the strongest BMP-2 secretion rates in osteoblastically-differentiated MSCs. Regardless of the promoter driving the transgene, a plateau of maximal BMP-2 secretion persisted for at least 21 d after the hBMP-2 gene electrotransfer. The present study demonstrates the feasibility of gene electrotransfer for efficie...

Reprogramming of mesenchymal stem cells derived from iPSCs seeded on biofunctionalized calcium phosphate scaffold for bone engineering

“…The BMP2 protein secretion from iPSC-MSCs and BMP2-iPSC-MSCs (passage 5 to 8) was evaluated by measuring BMP2 amount in the conditioned medium, as described previously [23]. Briefly, cells were cultured on 75 cm 2 TCPS flasks and the cell supernatants were collected during a 24-hour culture.…”

“…Gene therapy strategy has been used for this purpose because it has advantages over direct growth factor delivery, such as low cost and sustained long-term release [20]. Bone morphogenic protein 2 (BMP2), a member of the transforming growth factor β (TGFβ) superfamily, has a remarkable capability of inducing bone formation [21–23]. Preclinical and clinical investigations have successfully applied BMP2 in the therapy of bone defects [24].…”

“…Preclinical and clinical investigations have successfully applied BMP2 in the therapy of bone defects [24]. Various vectors have been tested to deliver BMP2 by gene therapy, such as liposome-mediated plasmid DNA delivery, adenoviral vectors, and lentiviral vectors [21–23]. Lentiviral vectors with minimal immunogenicity and improved biosafety after a series of modifications could transduce BM-MSCs with high efficiency and long-term stability, which were used to repair bone defect with superior mechanical quality [22,25,26].…”

Bone formation in rabbit’s leg muscle after autologous transplantation of bone marrow-derived mesenchymal stem cells expressing human bone morphogenic protein-2

“…The CMV promoter once was used as the standard promoter in most BMP transgene expression systems 40. However, Ferreira et al 41. have proved a fact that elongation factor-1α, (EF-1α), β-actin and GAPDH promoters are more efficient than the viral promoters such as CMV in driving gene expression, and provided the first in vitro evidence for a safe alternative to viral methods that permit efficient BMP-2 gene delivery and expression in MSCs.…”

“…Transgene electrotransfer is first highly efficient nonviral gene transfer method for most primary cells and for hard-to-transfect cell lines; it looks like a promising method. Ferreira et al 41. have compared a BMP-2 secretion rate of the control under CMV, eIF4A1, EF-1α, β-actin, GAPDH, fibronectin or osteocalcin promoters using electrotransfer into rat MSCs and showed that BMP-2 production correlated with sustained (up to 21 d) hBMP-2 mRNA expression.…”