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Flowcytometry for Estimating Efficient Transfection of Bone Marrow Cells with BCR-ABL Gene

Authors: Rihab Nasr --- Manal Adnan Habib --- Ahmad Iskandarani
Journal: Iraqi Academic Scientific Journal المجلة العراقية للاختصاصات الطبية ISSN: 16088360 Year: 2012 Volume: 11 Issue: supplement Pages: 649-653
Publisher: The Iraqi Borad for Medical Specialization المجلس العراقي للاختصاصات الطبية


ABSTRACT:BACKGROUND:Chronic myeloid leukaemia can be accurately modeled in laboratory mice by the retroviral transfer of a BCR-ABL gene into murine hematopoietic stem and progenitor cells, followed by transplantation of these cells into irradiated recipient mice.OBJECTIVE:The use of Retroviral vector for transfer of BCR-ABL gene into murine bone marrow cells (BMC) and measurement of efficiency of transfection by flowcytometry.METHODS:Murine bone marrow cells obtained from mice were cultured in a medium containing the supernatant of BCR-ABL (p210) transfected platinum E cells which is rich in retroviral vector carrying the BCR-ABL (p210) gene.The vector express green fluorescent protein (GFP) as well so that the efficiency of transfection of murine BMC with the target gene was able to be measured using flowcytometry.RESULTS:The use of the retrovirus packaging cell line enhanced the transduction of BMC with the retroviral vector and efficiency of transfection was 72% as measured by the flowcytometry.CONCLUSION:Transfer of BCR-ABL gene into murine BMC by retroviral vector that carry GFP marker which allowed the estimation of transfection efficiency by the flowcytomery

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