Cell Cultivation Laboratories

The cell culture laboratory has two rooms, one located in building 26 and another in building 29  of the CCQFA, UFPel, being coordinated by professors Roselia Maria Spanevello and Francieli Moro Stefanello in collaboration with the researcher Elizandra Braganhol (collaborator of PPGBBio). Cell culture laboratories are specialized in performing primary cell culture and in culturing cell lineages. Primary cultures performed include mouse peritoneal macrophage culture and neonatal rat brain cortical astrocytes. In addition, the laboratory also cultivates and maintains different cell lineages, some of them representative of normal cells, such as fibroblast (3T3-NHI), monocytes (THP-1) and microglia (N9) lineages, as well as representative lineages of breast tumors (MCF-7), prostate (DU145), skin (B16F10; A379) and central nervous system (C6, GL261, U251MG, U87MG). Among the techniques routinely performed in the laboratory are cell viability analysis tests using the MTT [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide], SRB (sulfarodamine-B) and LDH (lactate dehydrogenase); cell cycle progression analysis assays (labeling with propidium iodide), determination of cell death (double labeling with propidium iodide/annexin-V) and expression of membrane markers (labeling with specific antibodies) using flow cytometry and evaluation of cellular morphology change in control situations and after treatment through image capture using inverted microscope and immunocytochemistry techniques.

Infrastructure and Equipment: The cell culture laboratory has two air-conditioned rooms containing two laminar flow hoods, two CO2 incubators for cell culture, two water baths, two optical microscopes, an inverted microscope, a nitrogen barrel, two refrigerators duplex refrigerator, a vacuum pump, an analytical balance, a pH meter, a magnetic stirrer, two vortex apparatus and a centrifuge. For the preparation of material the laboratory has a water distiller, an autoclave and an air circulation oven for drying material.