Enrichment of cancer stem-like cells in glioblastoma multiforme

• Grant number: 14/26637-8
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: March 01, 2015
• End date: December 31, 2015
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Alexander Henning Ulrich
• Grantee: Maria Elisa Almeida Góes
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 12/50880-4 - Stem cells: from basic studies of kinin and purinergic receptor roles towards therapeutical applications, AP.TEM

Abstract

Glioblastoma (GBM) stem-like cells (GSC) represent the most undifferentiated and aggressive state of GBM malignant cells. GSC are postulated to be resistant against current therapy and able to self-renew and differentiate into the bulk cells of the tumour, playing a relevant role in GBM carcinogenesis and reoccurrence. Since its first experimental evidences, the enrichment of GSC in vitro has been widely performed. However, the methods pursued to achieve this enrichment have been poorly explored. In this way, the aim of this project is to evaluate different substrates for GSC culture and enrichment. For this, a GBM cell line will be cultured using four different substrates, with two being adequate for suspension cultures and two for monolayer cultures. Following this, GSC will be characterized regarding the expression of stemness markers CD133, Nestin, Sox-2 and ALDH1 by flow cytometry and their in vitro invasion pattern by matrigel invasion assay. In parallel, primary GBM cultures will be established in our laboratory, aiming to validate the enrichment of GSC based on the GBM cell line. This project attempts to better elucidate the biology of GSC and also to propose a protocol for in vitro GSC enrichment, thus supporting future study of this distinct GBM cells subpopulation.