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Articles by Jason Letourneau in JoVE
Ensaio in vitro da adesão bacteriana em mamíferos Células Epiteliais
Jason Letourneau, Cynthia Levesque, Frederic Berthiaume, Mario Jacques, Michael Mourez
Universite de Montreal, Groupe de Recherche sur les Maladies Infectieuses du Porc GREMIP, Faculte de medecine veterinaire
Este protocolo é um ensaio de adesão bacteriana simples que consiste na contagem do número de unidades formadoras de colônia de bactérias que são aderidos em células em cultura. O ensaio é robusto, independente da adesina estudado, e inúmeras variações são usadas na maioria dos laboratórios que trabalham com patogênese bacteriana.
Other articles by Jason Letourneau on PubMed
Phenotypic Characterization and Functional Analysis of Human Tumor Immune Infiltration After Mechanical and Enzymatic Disaggregation
Journal of Immunological Methods. Sep, 2011 | Pubmed ID: 21782822
Multi-parametric flow cytometry analysis is a reliable method for phenotypic and functional characterization of tumor infiltrating immune cells (TIIC). The isolation of infiltrating leukocytes from solid tumors can be achieved through various methods which can be both enzymatic and mechanical; however, these methods may alter cell biology. The aim of this study was to compare the effects of three tissue disaggregation techniques on TIIC biology in breast, kidney and lung tumor specimens. We therefore compared two enzymatic treatments using either collagenase type IA alone or in combination with collagenase type IV and DNase I type II, and one mechanical system (Medimachine™). We evaluated the impact of treatments on cell viability, surface marker integrity and proliferative capacity. We show that cell viability was not significantly altered by treatments. However, enzymatic treatments decreased cell proliferation; specifically collagenases and DNase provoked a significant decrease in detection of surface markers such as CD4, CD8, CD45RA and CD14, indicating that results of phenotypic studies employing these techniques could be affected. In conclusion, mechanical tissue disaggregation by Medimachine™ appears to be optimal to maintain phenotypic and functional TIIC features.