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.