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Chianget, G.G., Rubin, H.I., Cherington, V., Wang, T., Sobolewski, J., McGrath, C.A., Gaffney, A., Emami, S., Sarver, N., Levine, P.H., Greenberger, J.S. and Hurwitz, D.R. (1999) Bone marrow stromal cell-mediated gene therapy for hemophilia A, in vitro expression of human Factor VIII with high biological activity requires the inclusion of the proteolytic site at amino acid 1648. Human Gene Therapy, 10, 61-76. http://dx.doi.org/10.1089/10430349950019192

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• TITLE: Plasmid vector based generation of transgenic mesenchymal stem cells with stable expression of reporter gene in caprine

  AUTHORS: Manish Kumar, Renu Singh, Kuldeep Kumar, Pranjali Agarwal, Puspendra Saswat Mahapatra, Abhisek Kumar Saxena, Ajay Kumar, Subrata Kumar Bhanja, Dhruba Malakar, Rajendra Singh, Bikas C. Das, Sadhan Bag

  KEYWORDS: Transgenic MSC; Caprine; Plasmid Vector; Characterisation; In Vitro Differentiation

  JOURNAL NAME: Stem Cell Discovery, Vol.3 No.4, October 30, 2013

  ABSTRACT: The production of cells capable of expressing gene(s) of interest is important for a variety of applications in biomedicine and biotechnology, including gene therapy and a novel method of stem cell therapy in the various diseases. Achieving high levels of transgene expression for the longer period of time, without adversely affecting cell viability and differentiation capacity of the cells, is crucial. In the present study, we investigated the efficiency of plasmid vector for the production of transgenic cMSCs and examined any functional change of cells after transfection. To do so first we have collected bone marrows from the adult goats and cultured them for isolation of mesenchymal stem cells (cBM-MSCs). These cells were characterized using MSC specific markers including differentiation into osteocytes and adipocytes. Transfection with plasmid vector did not adversely affect cBM-MSCs morphology, viability or differentiation potential, and transgene expression levels were unaffected beyond passage 12th. The results indicated that we have been able to generate transgenic caprine MSC (tcBM-MSC) and transfection of cBM-MSCs using plasmid vector resulted in very high and stable transfection efficiency. This finding may have considerable significance in improving the efficacy of MSC-based therapies and their tracking in animal model.