Increased Viability by Over-Expression of Bcl-XL in a Human Neural Progenitor Cell Line.

Ahn, Young Hwan; Shin, Yong Sam; Yoon, Soo Han; Cho, Ki Hong; Cho, Kyung Gi; Kim, Seung Up

Case Report

Journal of Korean Neurosurgical Society 2004;35(4): 415-421.

Young Hwan Ahn, Yong Sam Shin, Soo Han Yoon, Ki Hong Cho, Kyung Gi Cho, Seung Up Kim

¹Department of Neurosurgery, Ajou University School of Medicine, Suwon, Korea. yhahn@ajou.ac.kr
²Department of Brain Disease Research Center, Ajou University School of Medicine, Suwon, Korea.

ABSTRACT

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.

Key Words: Bcl-XL; Human neural progenitor cell; Overexpression; Transfection