Osteogenic differentiation of mesenchymal stem cells by adenovirus mediated BMP-2 gene transfer

Matišić, Mirta (2013) Osteogenic differentiation of mesenchymal stem cells by adenovirus mediated BMP-2 gene transfer. Diploma thesis, Faculty of Science > Department of Biology.

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Human mesenchymal stem cells (hMSCs) are a unique population of adult stem cells which can be found in different organs and tissues in an adult organism, but they are originally isolated from bone marrow. Being multipotent, the hMSCs have a great capacity of differentiation in different tissues of mesenchymal origin, including bone, cartilage, muscle and adipose tissue. The hMSCs are greatly used in research of regenative medicine because of their simple isolation, fast multiplication in vitro and multipotency. In order to direct the hMSCs towards the osteogenic differentiation, a specific biological signal is needed to induce the differentiation. Such an osteoinductive signal is the bone morfogenetic protein 2 (BMP-2). Replication-defective adenovirus serotype 5 vector (RdAd5) which exhibits deletions in the E1 region, was used for gene transfer. Adenoviruses RdAd5 were multiplified in AD-293 cells, because they are able to replicate only in certain cells which have integrated E1 region. Research includes experiments which can be divided into two groups: the first group refers to multiplication, purification and determination of viral concentration and another one is associated with transduction of mesenchymal stem cells, osteogenic confirmation which is based on cytochemical staining and specific osteogenic markers. The main aims of this study are: virus cultivation and determination of virus titer, visualisation and determination of transduction level and confirmation of osteogenesis of mesenchymal stem cells.

Item Type: Thesis (Diploma thesis)
Keywords: mesenchymal stem cells, adenoviruses, BMP-2, osteogenesis
Supervisor: Marijanović, Inga
Co-supervisor: Matić, Igor
Date: 2013
Number of Pages: 45
Subjects: NATURAL SCIENCES > Biology
Divisions: Faculty of Science > Department of Biology
Depositing User: Grozdana Sirotic
Date Deposited: 20 Feb 2014 13:40
Last Modified: 22 Sep 2014 12:39
URI: http://digre.pmf.unizg.hr/id/eprint/458

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