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KMID : 1001920040350040415
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Journal of Korean Neurosurgical Society
2004 Volume.35 No. 4 p.415 ~ p.421
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Increased Viability by Over-Expression of Bcl-XL in a Human Neural Progenitor Cell Line
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Ahn Young-Hwan
Shin Yong-Sam
Yoon Soo-Han
Cho Ki-Hong
Cho Kyung-Gi
Kim Seung-Up
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Abstract
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Objective : This study is aimed whether bcl-xl could protect C4 cells from the cell death induced by serum deprivation.
Methods : The transient transfection of the bcl-xl gene was made with a LipofectAMINE reagent. An immunohistocytochemical assay and Western-blotting were performed to examine the bcl-xl transfection into the C4 cells. In order to analyze.- the effect of the bcl-xl transfection, the number of cells on the well plate were serially counted each day, for 5 days, from the 2nd to the 6th day after transfection. The number of GFP-positive cells in the defined fields, following, serum deprivation, was counted using fluorescence microscopy, and the total number of viable cells, including transfected cells, were also assessed.
Results : Immunocytochemical staining showed positive cells in 52% of nestin staining, 60% of GFAP and 20% of MAP-2. The number of cells decreased after transfection using the LipofectAMINE in the serum free medium (p<0.001). Western blotting using an anti-human bcl-xl antibodies showed that bcl-xl was expressed in both the non-transfected and bcl-xl transfected C4 cells. Cell death in the C4 cells, and the number of cells, were serially monitored each day for 5 days. In the bcl-xl transfected cells, the cell death induced by serum deprivation was significantly inhibited and delayed compared to that in the control cells (p<0.001).
Conclusion : It is suggested that the bcl-xl transfected human neural progenitor cells might improve the survival of the grafted cells, and may be an alternative source of cells for neural transplantation in degenerative diseases.
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KEYWORD
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Bcl-XL, Human neural progenitor cell, Overexpression, Transfection
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