The Cell Biology Unit (UBC) is a technology-based platform whose mission is to offer advice and support to several research groups or industries in different projects that use cell cultures and flow cytometry as a main technology. Since 2007, our unit carries out collaborative actions with the Servicio de Clasificación Celular y Citometría de Flujo (SECIF) from Clemente Estable Institute.
Using these technologies (cell culture and flow cytometry) we develop research projects in the area of biotechnology.
The UBC is also member of the Molecular, Cellular and Animal Technology Program (ProTeMCA), one of the institutionally supported programs.
CELL TECHNOLOGY:
Our group has been dedicated to the generation and characterization of recombinant cell lines of biomedicine and biotechnological interest. Among them, we use a great variety of reporter cell lines (IFN type I, NF-kB, redox biosensors, among others) to search for and characterize substances that interfere with the signaling pathways of type I IFNs (Burgi et al, 2012 and Burgi et al, 2016). Also we use them in in-vitro models of inflammation (NF-kB, Tiscornia et al, 2012 Mastropietro et al, 2015; Rolny et al, 2016) or to improve the metabolism/productivity of cells of biotechnological interest (redox biosensors within the framework of ProTeMCA).ENVIRONMENTAL TOXICOLOGY:
Endocrine disruptors (ED) are anthropogenic substances present in the environment, capable of altering the homeostasis of the endocrine system of
organisms, contributing to the development of pathologies. Our working hypothesis is that the increase of certain reproductive pathologies is caused, in part, by the increasing exposure to EDs present in the environment. In this context, we have validated an in vivo model —the transgenic mouse Oct4-GFP— for toxicological monitoring of environmental estrogens (Porro et al, 2015). We have also developed an in vitro assay that uses a dual reporter cell line and allows us to evaluate in a single test the estrogenic or androgenic activity of a putative ED. In this line of research, we are collaborating with Dr. Rodríguez (ISAL, CONICET-UNL, Argentina).ANTITUMORAL PEPTIDES:
Since 2011, we have collaborated with Dr. Vallespi, from the Center for Genetic Engineering and Biotechnology (CIGB), in Cuba, on the project “CIGB-552: novel peptide with antitumor and anti-inflammatory properties useful for cancer treatment”.
We demonstrate that CIGB-552 is effective in reducing the size of tumours present in mice and we identify the COMMD1 protein as a key mediator for its antitumor activity (Fernández Massó et al, 2013, Vallespí et al., 2014, Núñez de Villavicencio et al., 2015).
Recently, we described the minimum functional unit of CIGB-552 necessary to exercise its biological activity (ability to penetrate tumor cells, interact with COMMD1 and induce apoptosis, Astrada et al, 2016 and 2018).TISSUE ENGINEERING: INTESTINAL ORGANOIDS AND 3D CROPS:
Traditional cell cultures, in two dimensions, offer a simple, rapid, economic and reproducible evaluation system. However, they lack the ability to recreate the cellular interactions that take place in a tissue in vivo, limiting its predictive power. Our goal is to generate and characterize cellular models in three dimensions (3D) that resemble living systems, improving the correlation of results. In particular, we propose to establish a murine intestinal organoid culture, either from primary tissue or by differentiation of induced pluripotent stem cells (from the English iPSC) and apply them in the study of inflammation, cancer and probiotic screening.
LABORATORY OF CELLULAR CROPS
We have several laboratories equipped to work in an aseptic environment, such as biological safety cabinets, CO2incubators, cytological centrifuges and inverted microscopes.
We also have an Automatic Analyzer for Glucose and Lactate Bioprofile Basic 2 Analyzer (Nova Biomedical)
FLOW CYTOMETRY LABORATORY
Cellular analyzers:
BD FACSAria™ Fusion (BD Biosciences)
High speed cellular classifierSoftware for data analysis:
– Summit V4.3
– BD Accuri C6 Software V1.0.264.21
– FACSDiva
– FlowJo
We organize an annual training course for new users of the installed equipment.
We are open to establish collaborations and to participate in research projects providing technical and scientific assistance.For more information and making reservations, please contact us at: citometria@pasteur.edu.uy
Culture, amplification and storage of cell lines. We have a cellular bank with more than 20 certified cell lines (ATCC, DSMZ, Riken).
Detection of contamination by Mycoplasma sp in cell cultures using PCR.
Quantification of glucose and lactate in cell culture supernatants.
Cell-based assays: cytotoxicity, proliferation, transfection, generation of stable cell lines.
Experimental design, sample processing, acquisition and analysis by flow cytometry.
Training and advice for cytometry users.
Isolation of different cell populations from a mixture by high-speed cell separation (up to 4 pathways).
Cellular cloning by depositing a single cell per well using a cell sorter.
Analysis by Flow Cytometry: Procedure and Costs
Request form for analysis using Analytical Cytometers-CyAn ADP / BD Accuri C6 / Attune NXT
Analysis request in BD FACSAria Fusion
Latin American School of Flow Cytometry. October 1-5, 2018. Institut Pasteur de Montevideo and Faculty of Medicine, Uruguay. Funded by ANII, IPMontevideo, GRCF, ISAC, Udelar, PEDECIBA, ProInBio. Course coordinators: Bollati M, Lens, Giordano,Blanco, Malusardi.
Course “Alternative Methods to Lab Animals use”.May 21-25, 2018. Institut Pasteur de Montevideo, Uruguay. Funded by IPMontevideo, Premasur, LÓreal. Course coordinators: Bollati M, Crispo M, De Vecchi R.
International Course “Cell and Animal Models for Drug Discovery”. October 16 to 27, 2017. Institut Pasteur de Montevideo. Funded by ICGEB, RIIP, UNU BIOLAC, ESACT and FOCEM. Course coordinators: Bollati M, Crispo M, Comini M, Alves P.
Basic course in flow cytometry and its applications in research. February 22-26, 2016. Institut Pasteur de Montevideo, IIBCE, Uruguay. Funded by PEDECIBA. Coordinators of the course: Victoria S, Perelmuter K, Bollati M.
International course “Flow cytometry and cell sorting in biotechnology and biomedicine research”. April 17-28, 2014. Institut Pasteur de Montevideo. Funded by RIIP, ICGEB, UNU-Biolac and TWAS. Course coordinators: Bollati M, Gröbe L, García JM.
Course “Fundamentals and applications of flow cytometry”. October 7-18, 2013, Institut Pasteur de Montevideo, IIBCE, Uruguay. Sponsored by PEDECIBA and ProInBio.Course coordinators: Folle G, Porro V and Bollati M.
IV Latin American Seminar on Cellular Technology. November 7-9, 2010 – Hotel Ermitage, Montevideo. Funded by ICGEB, UNU-Biolac. Organizing Committee: Bollati M, Kratje R.
International course “Animal Cell Biotechnology: Product from cells, cells as product”, November 10-19, 2010, Institut Pasteur de Montevideo, Uruguay. Funded by ICGEB, UNU-Biolac. Organizing Committee: Bollati M, Kratje R.
International Course. MidTerm HEVAR Conference on Viral vectors as genetic vaccines against pathogens. April 7-13, 2008, University of the Republic – Institut Pasteur of Montevideo, Uruguay. Organizing Committee: Arbiza J, Epstein A, Fraefel C, Glikmann G, Bollati M.
2019-2021 – New molecular tools for reducing production costs of bioactive molecules. FMV_3_2018_1_148443. Responsible: Cecilia Abreu. Member of the research team: Mariela Bollati.
2019-2021 – Generation of polyphenol-rich extracts from grape pomaces. Alliance for Innovation – ANII ALI_2_2018_1_149574. Responsible: Margot Paulino. Member of the research team: Mariela Bollati.
2019-2020 – Cannabis and autism: characterization, extraction and effects in animal and cellular models. ALI_1_2018_1_147904. Responsible: Inés Carrera. Member of the research team: Mariela Bollati.
2018-2020 – Mitochondrial bioenergetics in senescence induced by melanoma therapy: assessing the impact on tumor development. FCE_1_2017_1_136021. Responsible: Celia Quijano. Consultant: Mariela Bollati.
2018-2019 – In silico approach for the identification and evaluation of mechanisms of action of bioactive peptides from different food sources. FSDA_1_2017_1_143964. Responsible: Margot Paulino. Member of the research team: Mariela Bollati.
2016-2018 – Generation and characterization of in vitro models for the study of endocrine disruptors. Responsible: M. Bollati. MOV_CO_2015_1_110054
2015-2018 – Design of biosensors for simultaneous monitoring of redox signalingand cAMP: From the computer to the cell and back to the computer. Responsible: S. Pantano. FMV_1_2014_1_104000
2014-2015 – High speed, multiparametric and biosecure cellular separator for biomedicine and biotechnological use. National Agency for Research and Innovation, ANII (EQC_2013_X_1_2, Uruguay). Principal Investigator: Mariela Bollati-Fogolin.
2013-2015 – Risk assessment for exposure to anthropogenic environmental estrogens in a model of transgenic mice Oct-4-GFP. National Agency for Research and Innovation, ANII. María Viñas Fund PR_ FMV_2_ 2011_1_6046 (Uruguay). Principal Investigator: Mariela Bollati-Fogolin
2012-2013 – Monitoring of natural and synthetic compounds that modulate the biological activity of type I interferons using a new reporter gene assay. Bilateral international cooperation Uruguay-Argentina (DICyT and MEC), Researchers Responsible: Mariela Bollati-Fogolín (Uruguay), Marcos Oggero (Argentina)
Relationship with the productive sector
The UBC has been working with the following companies by provision a particular
service, on a technological development contract, and as a consultant:
Mega Pharma (Uruguay): 2018
Biogénesis Bagó (Argentina): 2015-2019
BIOPOLIS (España), 2009-2013.
MICROSULES Laboratories (Uruguay), 2012-2013.
GRANUY (GRANAR GROUP, Paraguay – Uruguay), 2012-2013.
VIRBAC GROUP (France), 2011.
Laboratories SANTA ELENA SA (Uruguay), 2010–2011.
DANONE Research (France), 2007-2009.
CLAUSEN Laboratories(Uruguay), 2007–2018.
