EVALUATION OF THE EFFICIENCY OF PIRES2-EGFP AND PCDNA3-HBDNF-V5 PLASMIDS IN TRANSFECTION OF CCE ES CELLS BY THE ELECTROPORATION METHOD

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FATHI F.; ALTARIHI T.; MOVAHEDIN M.; MOULA SEYED JAVAD

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Journal Paper

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Journal: Cell Journal (Yakhteh) Year:2006 | Volume:8 | Issue:1 (29) Page(s): 17-22

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Title

EVALUATION OF THE EFFICIENCY OF PIRES2-EGFP AND PCDNA3-HBDNF-V5 PLASMIDS IN TRANSFECTION OF CCE ES CELLS BY THE ELECTROPORATION METHOD

Author(s)

FATHI F. | ALTARIHI T. | MOVAHEDIN M. | MOULA SEYED JAVAD | Issue Writer Certificate

Keywords

EMBRYONIC STEM CELLS (ESCS)Q2

TRANSFECTIONQ2

ELECTROPORATIONQ2

GFPQ2

BDNFQ2

Abstract

Introduction: Unlimited self renewal and potential capacity of embryonic stem cells (ESCs) in differentiating into a wide variety of cell types has made the cells an attractive source of donor cells for developmental studies and cell therapy. The aim of the present study was the evaluation of the TRANSFECTION efficiency of pIRES2-EGFP and pcDNA3-hBDNF-v5 plasmids in CCE ES cells by the ELECTROPORATION method. Material and Methods: The plasmids transformed into DH5α competent bacteria and propagated as maxi-prep. The plasmids then purified and transfected into ESCs by means of the ELECTROPORATION method. To confirm the expression of the GFP and BDNF genes, invert fluorescent microscopy and RT-PCR were used. Expression of EGFP was confirmed by examining the transfected cells with fluorescent microscopy. In case of BDNF, total RNA extracted from stably transfected cells, were quantitatively evaluated by spectrophotometry and qualitatively by agarose gel electrophoresis. Then mRNAs were reverse transcripted and BDNF cDNA amplified by specific primers.Results: The products of the PCR were separated and visualized on agarose gel electrophoresis. Both techniques revealed a successful TRANSFECTION of CCE ES cells by both plasmids.Conclusion: The obtained data indicated that the CMV promoter is active in undifferentiated mouse ESCs and a CCE cell line is an appropriate donor cell for cell-mediated BDNF gene transfer.

Cites

OPTIMIZING A LIPOCOMPLEX-BASED GENE TRANSFER METHOD INTO HELA CELL LINE (SHORT COMMUNICATION)

References

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APA: Copy

FATHI, F., ALTARIHI, T., MOVAHEDIN, M., & MOULA, SEYED JAVAD. (2006). EVALUATION OF THE EFFICIENCY OF PIRES2-EGFP AND PCDNA3-HBDNF-V5 PLASMIDS IN TRANSFECTION OF CCE ES CELLS BY THE ELECTROPORATION METHOD. CELL JOURNAL (YAKHTEH), 8(1 (29)), 17-22. SID. https://sid.ir/paper/381891/en

Vancouver: Copy

FATHI F., ALTARIHI T., MOVAHEDIN M., MOULA SEYED JAVAD. EVALUATION OF THE EFFICIENCY OF PIRES2-EGFP AND PCDNA3-HBDNF-V5 PLASMIDS IN TRANSFECTION OF CCE ES CELLS BY THE ELECTROPORATION METHOD. CELL JOURNAL (YAKHTEH)[Internet]. 2006;8(1 (29)):17-22. Available from: https://sid.ir/paper/381891/en

IEEE: Copy

F. FATHI, T. ALTARIHI, M. MOVAHEDIN, and SEYED JAVAD MOULA, “EVALUATION OF THE EFFICIENCY OF PIRES2-EGFP AND PCDNA3-HBDNF-V5 PLASMIDS IN TRANSFECTION OF CCE ES CELLS BY THE ELECTROPORATION METHOD,” CELL JOURNAL (YAKHTEH), vol. 8, no. 1 (29), pp. 17–22, 2006, [Online]. Available: https://sid.ir/paper/381891/en

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