Analysis of methodological techniques for optimization of in vitro bioluminescence studies

• Grant number: 17/16981-1
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: January 01, 2018
• End date: December 31, 2018
• Field of knowledge: Biological Sciences - Genetics - Human and Medical Genetics
• Principal Investigator: Nelson Gaspar Dip Júnior
• Grantee: Luis Felipe Gastaldo Poletti
• Host Institution: Hospital do Servidor Público Municipal (HSPM). Secretaria Municipal de Saúde (São Paulo). São Paulo , SP, Brazil

Abstract

Bioluminescence is the ability of certain organisms to generate their own light from specific enzymatic reactions. Studies using the bioluminescence process have set a major milestone in science, since the technique has made it possible to visualize intracellular metabolic processes in a practical and easy-to-reproduce way. Its application allowed the analysis of cellular processes such as cancer investigations, including molecular pathways of carcinogenesis, treatments efficiency and molecular target therapies, making it an important tool in the preclinical trials However, there are still standardizations to be made regarding its methodology, in order to improve the technique and therefore increase its effectiveness. Among the limiting factors of the technique the present study aims to investigate the processes of selection of stable cultures, in order to balance the desirable effects of the undesirable ones. Thus, we will investigate the following observed variables as limiting: To assess the impact of different puromycin concentrations on the cell viability of human bladder urothelial carcinoma 5637 tumor cell lines in order to determine the optimal antibiotic concentration for cell selection. To study the role of three types of lipid reagents in the transfection of the plasmid bioluminescence vector into human bladder urothelial carcinoma 5637 cells in an attempt to establish a standard method of plasmid transfection. (AU)