Repository of Research and Investigative Information

Repository of Research and Investigative Information

Kurdistan University of Medical Sciences

Genetic modification of bone-marrow mesenchymal stem cells and hematopoietic cells with human coagulation factor IX-expressing plasmids

(2016) Genetic modification of bone-marrow mesenchymal stem cells and hematopoietic cells with human coagulation factor IX-expressing plasmids. Biologicals.

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Ex-vivo gene therapy of hemophilias requires suitable bioreactors for secretion of hFIX into the circulation and stem cells hold great potentials in this regard. Viral vectors are widely manipulated and used to transfer hFIX gene into stem cells. However, little attention has been paid to the manipulation of hFIX transgene itself. Concurrently, the efficacy of such a therapeutic approach depends on determination of which vectors give maximal transgene expression. With this in mind, TF-1 (primary hematopoietic lineage) and rat-bone marrow mesenchymal stem cells (BMSCs) were transfected with five hFIX-expressing plasmids containing different combinations of two human β-globin (hBG) introns inside the hFIX-cDNA and Kozak element and hFIX expression was evaluated by different methods.In BMSCs and TF-1 cells, the highest hFIX level was obtained from the intron-less and hBG intron-I,II containing plasmids respectively. The highest hFIX activity was obtained from the cells that carrying the hBG intron-I,II containing plasmids. BMSCs were able to produce higher hFIX by 1.4 to 4.7-fold increase with activity by 2.4 to 4.4-fold increase compared to TF-1 cells transfected with the same constructs. BMSCs and TF-1 cells could be effectively bioengineered without the use of viral vectors and hFIX minigene containing hBG introns could represent a particular interest in stem cell-based gene therapy of hemophilias. © 2016 International Alliance for Biological Standardization.

Item Type: Article
Keywords: blood clotting factor 9; complementary DNA; hemoglobin beta chain; blood clotting factor 9; RNA directed DNA polymerase, animal cell; animal tissue; Article; bioengineering; bone marrow derived mesenchymal stem cell; cell lineage; controlled study; DNA modification; female; gene expression; genetic transfection; hematopoietic stem cell; hemophilia; hFIX gene; intron; nonhuman; plasmid; priority journal; protein synthesis; rat; stem cell gene therapy; animal; bone marrow cell; cell culture; gene therapy; gene vector; genetics; hematopoietic stem cell; hemophilia A; human; mesenchymal stroma cell; metabolism; procedures; tumor cell line, Animals; beta-Globins; Bone Marrow Cells; Cell Line, Tumor; Cells, Cultured; Factor IX; Gene Expression; Genetic Therapy; Genetic Vectors; Hematopoietic Stem Cells; Hemophilia A; Humans; Introns; Mesenchymal Stromal Cells; Plasmids; Rats; RNA-Directed DNA Polymerase; Transfection
Page Range: pp. 170-177
Journal or Publication Title: Biologicals
Volume: 44
Number: 3
Publisher: Academic Press
Identification Number: 10.1016/j.biologicals.2016.01.002
ISSN: 10451056
Depositing User: مهندس جمال محمودپور

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