EXPRESSION OF RECOMBINANT COAGULATION FACTOR IX IN HUMAN AMNIOTIC MEMBRANE-DERIVED MESENCHYMAL STEM CELLS: A NEW STRATEGY TO GENE THERAPY OF HEMOPHILIA B

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PARIDAR M.; AMIRIZADEH N.; HABIBI ROUDKENAR M.; AMIRI F.; ABOLGHASEMI H.; JALILI M.A.

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Journal Paper

Paper Information

Journal: Iranian Journal of Blood and Cancer Year:2014 | Volume:6 | Issue:3 Page(s): 133-141

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Title

EXPRESSION OF RECOMBINANT COAGULATION FACTOR IX IN HUMAN AMNIOTIC MEMBRANE-DERIVED MESENCHYMAL STEM CELLS: A NEW STRATEGY TO GENE THERAPY OF HEMOPHILIA B

Author(s)

Keywords

HEMOPHILIA B

AMNION MEMBRANE

MESENCHYMAL STEM CELL

FACTOR IX

GENE THERAPY

Abstract

Background: HEMOPHILIA B is an X-linked hereditary disorder of blood coagulation system which is caused by FACTOR IX (FIX) deficiency. FACTOR IX is a plasma glycoprotein that participates in the coagulation process leading to the generation of fibrin. Replacement of FACTOR IX with plasma-derived or recombinant FACTOR IX is the conventional treatment for HEMOPHILIA B to raise the FACTOR IX level to therapeutic range. Recently, GENE THERAPY has been regarded as a promising approach to treat HEMOPHILIA B. This study was aimed to express the FACTOR IX in human amniotic membrane-derived MESENCHYMAL STEM CELLs (hAM-MSCs).Materials and Methods: Human amniotic membrane-derived MESENCHYMAL STEM CELLs were isolated and characterized from AMNION MEMBRANE. FACTOR IX from commercially available plasmid was sub-cloned into pcDNA3.1 vector. Recombinant pcDNA3.1-FIX construct was confirmed by PCR, enzymatic digestion and DNA sequencing. MESENCHYMAL STEM CELLs were transfected with the recombinant vector. Expression of FACTOR IX was determined by RT-PCR, ELISA and its biological activity assay was performed using aPTT.Results: Isolated hAM-MSCs expressed specific MESENCHYMAL STEM CELLs markers and were able to differentiate to osteocytes and adipocytes lineages. hAM-MSCs expressed hrFIX at mRNA and protein level. The maximum amount of hrFIX was 120 ng/ml at 72 hrs after hAM-MSCs transfection. This hrFIX was biologically active (11% activity), formed fibrin clot in aPTT test and caused more than two fold decrease in clotting time.Conclusion: The hAM-MSCs expressing FACTOR IX would be useful for GENE THERAPY of HEMOPHILIA B. However further studies are required to prove these finding.

Cites

THE EFFICIENCY OF SECRETION AND γ-CARBOXYLATION OF RECOMBINANT HUMAN FACTOR IX IN STABLE DROSOPHILA CELLS

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APA: Copy

PARIDAR, M., AMIRIZADEH, N., HABIBI ROUDKENAR, M., AMIRI, F., ABOLGHASEMI, H., & JALILI, M.A.. (2014). EXPRESSION OF RECOMBINANT COAGULATION FACTOR IX IN HUMAN AMNIOTIC MEMBRANE-DERIVED MESENCHYMAL STEM CELLS: A NEW STRATEGY TO GENE THERAPY OF HEMOPHILIA B. IRANIAN JOURNAL OF BLOOD AND CANCER (IJBC), 6(3), 133-141. SID. https://sid.ir/paper/313178/en

Vancouver: Copy

PARIDAR M., AMIRIZADEH N., HABIBI ROUDKENAR M., AMIRI F., ABOLGHASEMI H., JALILI M.A.. EXPRESSION OF RECOMBINANT COAGULATION FACTOR IX IN HUMAN AMNIOTIC MEMBRANE-DERIVED MESENCHYMAL STEM CELLS: A NEW STRATEGY TO GENE THERAPY OF HEMOPHILIA B. IRANIAN JOURNAL OF BLOOD AND CANCER (IJBC)[Internet]. 2014;6(3):133-141. Available from: https://sid.ir/paper/313178/en

IEEE: Copy

M. PARIDAR, N. AMIRIZADEH, M. HABIBI ROUDKENAR, F. AMIRI, H. ABOLGHASEMI, and M.A. JALILI, “EXPRESSION OF RECOMBINANT COAGULATION FACTOR IX IN HUMAN AMNIOTIC MEMBRANE-DERIVED MESENCHYMAL STEM CELLS: A NEW STRATEGY TO GENE THERAPY OF HEMOPHILIA B,” IRANIAN JOURNAL OF BLOOD AND CANCER (IJBC), vol. 6, no. 3, pp. 133–141, 2014, [Online]. Available: https://sid.ir/paper/313178/en

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