Different Enzyme Cocktail Conditions Successfully Establish Primary Brain Tumor Cell Line Derived from Pilomyxoid Astrocytoma Patient

Abstract

Background and Objective : Pediatric brain tumor is the second most common cancer after leukemia. Multidrug resistance is a problematic issue worldwide resulting in poor prognosis. Molecular and biological research on cancer cells is essentially required to understand the nature of cancer cells but the most effectiveness of different enzyme cocktail conditions for primary cell line establishment has not been conclusive. This study tests the hypothesis that different enzyme cocktail conditions differently affect the quality of cell line establishment.  

Materials and Methods: Primary cell line establishment of pilomyxoid astrocytoma tissues was performed using 7 different enzyme cocktail conditions (C: collagenase, H: hyaluronidase, D: DNase I, C+D: collagenase + DNase I, C+H: collagenase + hyaluronidase, D+H: DNase I + hyaluronidase, and C+D+H: collagenase + DNase I + hyaluronidase) for cell dissociation. To compare the effectiveness of 7 enzyme cocktail conditions, a degree of cell dissociation, clump cell size, percentage of live cells, cell proliferation rate and time of 80% cell confluency from each condition were investigated.

Results: All of the 7 enzyme cocktail conditions provided similarly effective cell dissociation. All conditions displayed a high percentage of live cells after dissociation, ranging from 80-95%. Although, most of studied conditions could proliferate to reach 1.5x10⁵ cells within 96 hours after seeding. D, H and C+D conditions could not proliferate to reach the expected number within 120 hours. However, all conditions effectively grew to produce 80% cell confluency within 120 hours.

Conclusion: The present data indicate that in primary cell line establishment of brain pilomyxoid astrocytoma, different standard enzyme cocktail conditions similarly provide the effectiveness of cell line establishment.