“Simple Methods to Derive Primary Malignant Glioma Cell Lines and Assay of Cellular Damage for Preclinical Studies”

ABSTRACT

Primary malignant glioma cell lines are being used for initial screening of anticancer agents. We utilized a simple mechanical disaggregation method for deriving cell lines from tumor tissues; and a Coverslip Culture-Acridine Orange Staining method to study cellular damage. Cell lines could be grown for up to three passages within three weeks after surgery. Cell proliferation, total cellular damage, and MTT assay were studied as parameters of cytotoxic response. Frequencies of damaged cells varied in different cell lines; and increased after cytotoxic treatments under clinically relevant conditions. These methods could contribute to preclinical evaluation of treatment response before commencement of radio-chemotherapy.

KEYWORDS:

• Malignant gliomas
• Primary cell lines
• Radiation therapy
• Temozolomide
• 2-deoxy-d-glucose

Declaration of interest

The authors report no declarations of interest. This work was supported by Department of Biotechnology, Government of India [Grant No. BT/PR12932/MED/30/236/2009].

Acknowledgments

This work was supported by Department of Biotechnology, Government of India [Grant No. BT/PR12932/MED/30/236/2009]. Department of Neurosurgery, NIMHANS provided the tumor samples and U251MG cell line was kindly provided by Dr. Nandakumar, Department of Neurochemistry, NIMHANS, Bangalore. Statistical analysis was carried out with the help of Department of Biostatistics, NIMHANS, Bangalore. We gratefully acknowledge Dr. C. K. Chandrasekhar Sagar, Dr. Vani Santosh, Dr. Phalguni Alladi, Dr. T. R. Raju, Dr. Manjunatha Venkataswamy, Dr. S. K. Shankar and Dr. D. K. Subbakrishna for their valuable suggestions, discussions and support. Radiation facilities were provided by Radiation Therapy Department, Kidwai Memorial Institute of Oncology, Bangalore, India.

Additional information

Funding

Department of Biotechnology, Ministry of Science and Technology; [(BT/PR12932/MED/30/236/2009)]