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Inventory of the Spanish Institutions and Scientists Involved in Alternatives to the use of Laboratory Animals (Refinement, Reduction or Replacement)+

Inventory: 18h. Inventory of Institutions

Laboratory: Department of Cellular Biology. University of Barcelona
Avinguda Diagonal 645 - 08028 Barcelona Tel.: 93 402.15.50 Fax: 93 411.29.67
E-mail: celltec@bio.ub.es WEB: http://celltec.ub.es

Type of Institution: University.

Staff:

Senén Vilaró. E-mail: senen@porthos.bio.ub.es; General Manager. PhD. Professor of Cell Biology.
Manuel Reina. E-mail: mreina@porthos.bio.ub.es; Assistant Manager. PhD. Associate Professor of Cell Biology.
Ricardo Casaroli-Marano. E-mail: casarou@porthos.bio.ub.es; Project Manager. PhD. Assistant Professor of Cell Biology.
Roser Pagán. E-mail: pagan@porthos.bio.ub.es; Project Manager. PhD. researcher.
Myriam Fabre. E-mail: mfabre@porthos.bio.ub.es; Project Manager. PhD researcher.
Àlex Monnà. E-mail: amonna@fbg.ub.es; Promotion. Graduate in Biology.

In addition, there are 3 laboratory technicians and 13 post-graduates students, performing a staff with 22 people involved in alternative methods.

Activities/aims: This university department aims to promote innovation and transfer in the fields of cellular and molecular technology. It performs cell and molecular biology and toxicology testing on a routine basis, biokinetics and biotransformation, pathology, diagnostics and monitoring –chemical, biological- studies at a research level. Pharmaceuticals, cosmetics and diverse lead compounds are evaluated; is also interested in research related to in vitro models of different tissues for transplantation.

Model systems: the model systems used are in vitro methods –primary culture of endothelial cells of different species and cell lines of different endothelial cells; primary culture of smooth muscular cells from human aorta (migratory, quiescent, proliferating and confluent phenotypes); cell line of intestinal epithelial cells from human adenocarcinoma; primary culture of dermal fibroblasts from human skin; primary culture of cells from the pigmentary epithelium of the human retina; primary culture of human blood monocytes and cell line of human monocytes from human macrophages; cell line of kidney cells from dog; primary culture of queratinocytes from human skin; primary culture of hepatocytes (adult rat liver, regenerant liver from adult rat, liver from rat embrion and from human liver) and cell line hepatocytes from human hepatic cancer; neurones from human and rat brain; adipocytes from mouse adipose tissue and primary culture of adipocytes from rat and human adipose tissue; and other cellular models (human adrenal gland cells, immortalised human placenta cells, hamster ovary cells and different cell lines with constitutive and stable expression of receptors from the plasmatic membrane that are involved in the development of artheriosclerosis).

Experimental systems (examples):

Analysis of the flow of seed molecules through the blood brain barrier on cocultures systems involving endothelial cells and astrocytes as a model of the brain blood barrier.

Metabolic, functional and toxicological analysis of the effects of drugs on endothelial cells to study angiogenic molecules.

Analysis of the effect of seed molecules on artheriosclerotic processes on HASMC monolayer cultures as a model of artheriosclerosis patology.

System of functional screening of pharmacological and cosmetic active principles on cocultures of human keratinocytes and fibroblasts that mimic human skin.

Analysis of the functional effect of seed molecules driven to inflammatory, allergic, infectious and arteriosclerotic processes on cultures of Human Monocytes differentiated to macrophage.

In vitro analysis of renal absorption of seed molecules on polarised cultures of epithelial cellsa and on monolayer cultures of primary hepatocytes to study hepatotoxicity.

Analysis of seed molecules effects on differentiation, proliferation or neural toxicity on monolayer cultures of human or rat brain neurones.

Analysis of seed molecules on the accumulation of fat in adipocytes (monolayer cultures of rat, mouse or human adipocytes).

Work lines: It offers all biotechnological, pharmaceutical and cosmetics companies the experience and knowledge, concerning development and use of in vitro cellular models. The group has a wide experience in the screening and determination of efficacy, metabolism and toxicity of drugs, active principles and elaborated products. CELLTEC UB also offers to the R+D Departments of companies the possibility to obtain its advice and the transfer of its technology. It is developing different activities such as: Assays on in vitro cellular models. Research services: Development and validation of new in vitro cellular models; Research on mechanism of action, toxicity and metabolism of drugs and new molecules; Research on tissue bio-engineering; Cellular kit development and validation; Technology transfer; Scientific advice.

Quality assurance/Validation programmes: The team is available to participate in EU validation programmes and is in process to implement Good Laboratory Practices.

Sources of financing: National projects (CICYT, MINER, FEDER,CIRIT, public and private foundations, etc.), European projects (Frame Programmes) and contracts with industries (pharmaceutical, cosmetics, biotechnology, etc.). 

Staff:

Miguel Borrás Suárez, Email: jnadal@bio.ub.es, Dr Ciencias Biológicas.
Jacint Nadal Puigdefábregas, Email: jnadal@bio.ub.es, Dr Ciencias Biológicas.
Eulalia Delgado Sureda, Email: jnadal@bio.ub.es, Ld Ciencias Biológicas.

Total staff involved in alternative methods is 3 people.

Activities / aims: This university department is mainly involved in basic research; it performs toxicology (genotoxicity / mutagenicity, ecotoxicity) testing on a routine basis. pesticides and environmental pollutants are routinely evaluated.

Model systems: The model systems used are invertebrate animal models (earthworm)

The endpoint employed in vivo is single cell electrophoresis (COMET).

Experimental systems (examples):

1 earthworm coelomocytes, COMET assay, terrestrial ecotoxicity

Work lines: Effect of air pollution on individuals and populations of arthropods, mammals and birds. Biodiversity. Toxic markers include hematology, serum biochemistry, heavy metal analysis, histopathology; respiratory tract: light microscopy, transmission and scanning electron microscopy, microanalysis, image analysis. Soil pollution: single cell gel electrophoresis (Comet test) of vertebrate lymphocytes and earthworm coelomocytes.

Quality assurance / Validation programmes: A Quality Assurance Unit is operating inside the facility. The institution applies Good Laboratory Practices rules: implementation in course The team have previously been involved in the following validation programme of alternative methods: organochlorated determination. They are available to participate in EU validation programmes.

Sources of financing: Fuerzas Electricas de Cataluña (FECSA) / Fundació Bosch i Gimpera (1992-1995); Fundació La Caixa / Fundació Bosch i Gimpera (1993); Ministerio de Sanidad y Consumo (1994); Plan de Investigacón de Cataluña, GRQ 94-1050 (código del proyecto 02161) (1994); FECSA / Fundació Bosch i Gimpera, proyecto BG2392 (1995); Assesora DGICYT, proyecto PB94-0877 (1995); Plan de Investigación de Cataluña, 1996SGR 00072 (1996); Asesora DGICYT, proyecto PB96-0224 (1997).

Type of Institution: University

Staff:

Adela Mazo Sánchez, Email: adela@sgenz.bq.ub.es, Dr C Químicas, Profesor Titular de Bioquímica y Biología Molecular, 50 % time devoted to altern. meth., duties related to altern. meth.: Responsable Científico del Servicio de Cultivos Celulares de la Universidad de Barcelona
Manuel Cascalló Piqueras, Email: cpanc@sgenz.bq.ub.es, Ld Bioquímica, Becario, FPI Grupo de Ingeniería Celular y Terapia, 80 % time devoted to altern. meth.
Joaquín Calbó Angrill, Email: cpanc@sgenz.bq.ub.es, Ld Bioquímica, Becario, FPI Grupo de Ingeniería Celular y Terapia, 80 % time devoted to altern. meth.

Total staff involved in alternative methods is 3 people.

Activities / aims: This university department is mainly involved in basic research, non-regulated applied research and method development; it performs cell biology, molecular biology, toxicology (haematoxicity, genotoxicity, mutagenicity), diagnostics (genetic alterations in tumours) and culture methodology studies on a routine basis. Pharmaceuticals are routinely evaluated.

Model systems: The model systems used are conventional animal models (mice without thymus) and in vitro methods: cell lines culture (human pancreatic cell lines (NP9, NP18, NP29, NP31, etc)).

The endpoints employed are in vivo analysis of xenotransplant tumours treated with different protocols and in vitro morphology (optical and electronic microscopy), cell viability (trypan blue exclusion, MTT reduction) and cellular proliferation (cell count, WST test).

Experimental systems (examples):

1 human pancreatic cell lines: NP-9, NP-29, NP-31, NP-18 and sublines obtained by metastasis dissemination, propidium iodide by flow cytometry, housecat by microscopy and flow cytometry, anexin-V by fluorescence microscopy and flow cytometry, new therapeutic approaches by suppressor reintroduction.

Work lines: The focus of the research is addressed to the development of new therapeutical approaches based on the combination of genotoxic drugs and the reintroduction of tumour suppressor genes (p53, p16, etc) which potentiates drug action. It is used as a model in the human exocrine pancreatic cancer.

Quality assurance / Validation programmes: The team are available to participate in EU validation programmes in relation to diagnostic methods and cancer therapy.

Sources of financing: FIS, Maratón del Cáncer (TV3), Plan Nacional de Salud CICYT

Type of Institution: University

Staff:

Anna Mª Gómez Foix, Email: anamaria@sun.bq.ub.es, Dra Farmacia, Prof. Titular, 100 % time devoted to altern. meth.
Celia García Martínez, Email: celia@sun.bq.ub.es, Dr Biología, Investigador Contratado, 100 % time devoted to altern. meth.
Eulalia Montell Bonaventura, Email: laia@sun.bq.ub.es, Doctoranda, Beca FPI, 100 % time devoted to altern. meth.
Alexandra Arias Piñeiro, Email: sandra@sun.bq.ub.es, Técnico, Técnico Contratado, 100 % time devoted to altern. meth.
Josep Carles Jiménez Chillarón, Email: jimenez@sun.bq.ub.es, Ld Biología, Beca Generalitat Catalunya, 100 % time devoted to altern. meth.

Total staff involved in alternative methods is 5 people.

Activities / aims: This university department is mainly involved in basic research and alternatives to animals in education; it performs human biology, cell biology, molecular biology, biochemistry, nutrition, culture methodology studies on a routine basis. Hormones, biomaterials and nutrients are routinely evaluated.

The main use of alternative methods is for complementary and replacement studies.

Model systems: The model systems used are conventional animal models and transgenics, in vitro methods -- primary culture of dispersed cells (human muscle cells), cell lines culture (muscle C2C12), and education models (cell lines, muscle, kidney).

The endpoint employed is in vitro metabolic activity

Experimental systems (examples):

1 primary culture of human muscle cells, metabolism of glucose and lipids, metabolic regulation, replacement and complementary test

Work lines: Development of muscle gene therapy strategies for the treatment of diabetes and inborn myopathies. Viruses with target proteins are constructed and their impact is tested in a primary cultured human muscle model and in vivo rodent models.

Quality assurance / Validation programmes: The team are available to participate in EU validation programmes.

Sources of financing: CICYT, BIOMED2 Programme

Type of Institution: University

Staff:

José Domenech Berrozpe, Email: domenech@farmacia.far.ub.es, Dr Farmacia, Catedrático, 10 % time devoted to altern. meth.
Concepción Peraire Guitart, Email: mcper@farmacia.far.ub.es, Dr, Profesor Titular, 10 % time devoted to altern. meth.
Helena Colom Codina, Email: colom@farmacia.far.ub.es, Dr, Profesor Asociado, 10 % time devoted to altern. meth.
Ana Cristina Calpena Campmany, Email: calpena@farmacia.far.ub.es, Dr Farmacia, Titular de Universidad, 10 % time devoted to altern. meth.
Jacinto Lauroba Viladrosa, Email: jaclau@farmacia.far.ub.es, Dr Farmacia, Titular de Universidad, 10 % time devoted to altern. meth.
Elvira Escribano Ferrer, Email: elvira@farmacia.far.ub.es, Dr Farmacia, Titular de Universidad, 10 % time devoted to altern. meth.
Antonio Boix Montañés, Email: boix@farmacia.far.ub.es, Dr Farmacia, Prof. Asociado de Universidad, 10 % time devoted to altern. meth.
Ignacio Díez Martín, Dr Farmacia, Prof. Asociado de Universidad 10 % time devoted to altern. meth.

Total staff involved in alternative methods is 8 people.

Activities / aims: This university department is mainly involved in non-regulated applied research, method validation and alternatives to animals in education; it performs biokinetics, biotransformation and evaluation studies of pharmaceuticals by transdermal route testing on a routine basis. Pharmaceuticals and colourings are routinely evaluated; diverse chemical compounds also being occasionally studied.

Model systems: The model systems used are education models (flux) and mathematical modelling (exponential, linear and non-linear).

Work lines: Transdermal delivery systems, dissolution tests, PK-PD, evaluation of pharmaceutical dosage forms.

Quality assurance / Validation programmes: The team are available to participate in EU validation programmes.

Sources of financing: Proyectos de colaboración universidad/empresa (Fundació Bosch-Gimpera), Plan Nacional de I+D

Type of Institution: University

Staff:

Jorge Camarasa García, Email: camarasa@far.ub.es, Dr Farmacia, Catedrático de Farmacología, duties related to altern. meth.: Representante Español en Meeting sobre simulaciones en docencia
Elena Escubedo Rafa, Email: escubedo@far.ub.es, Dr Farmacia, Profesora Titular de Farmacología.

Total staff involved in alternative methods is 2 people.

Activities / aims: This university department is mainly involved in basic research and alternatives to animals in education; it performs pharmacodynamics, pharmacology (molecular pharmacology) and toxicology (neurotoxicity) testing on a routine basis, and molecular biology studies at a research level. Pharmaceuticals and toxins are routinely evaluated.

Model systems: The model systems used are conventional animal models , in vitro methods -- primary culture of dispersed cells (rat cerebellar granular cells) and education models (pharmacodynamics simulations).

The endpoints employed in vitro are cell viability and cytoskeleton/membranes/enzyme release.

Experimental systems (examples):

1 cerebellar granular cells, propidium iodide, rhodamine 123, neuronal physiology.

Work lines: Study of molecular and cellular aspects on neuronal cell death. Study of different biochemical cell markers, mainly heat shock protein expression, on neurodegerative animal models. Study of the interaction between microglial and lymphocyte cells that can led to neuronal death.

Quality assurance / Validation programmes: A Quality Assurance Unit is operating inside the facility. The institution applies Good Laboratory Practices rules according to the EU. The team are available to participate in EU validation programmes.

Sources of financing: CICYT, Universidad de Barcelona, private laboratories

Type of Institution: University

Staff:

M.Pilar Vinardell Martinez-Hidalgo, Email: pilarv@farmacia.far.ub.es, Dr Farmacia, Profesor Titular de Fisiologia, 100 % time devoted to altern. meth.
Montserrat Mitjans Arnal, Email: mitjans@farmacia.far.ub.es, Dr Farmacia, Profesor Asociado de Fisiologia, 50 % time devoted to altern. meth.

Total staff involved in alternative methods is 2 people.

Activities / aims: This university department is mainly involved in basic research, regulatory testing and alternatives to animals in education; it performs toxicology (ocular, dermal irritation and corrosivity) and cell biology and biochemistry stydies at a research level. Surfactants are routinaly evaluated; diverse chemical compounds also being occasionally studied.

Model systems: The model systems used are chicken embryo chorioalantoid membrane assay (HET-CAM), CAM-TBS Test, RBC test system, RBC photo assay-photohaemolysis and haemoglobin oxidation, haemoglobin denaturation test. The endpoints employed are lesions to blood vessels (CAM), erythrocyte haemolysis and haemoglobin denaturation. 

Experimental systems (examples):

1. Chick embryo chorioallantoid membrane assay, blood vessel lesions, ocular irritation, replacement test.
2. Erythrocyte haemolysis to quantify adverse effects of surfactants and detergent products on the cytoplasmic membrane in combination with the damage of liberated cellular proteins, ocular and dermal irritation replacement test.
3. Phototoxic potentials of chemicals by their ability to disturb the erythrocyte membrane under UV irradiation as a replacement test.

Work lines: The focus of the research is addressed to the development of new in vitro methods to investigate the potential ocular and dermal irritation of surfactants and chemicals.

Quality assurance / Validation programmes: The Quality Assurance Unit of the Barcelona University is operating inside the facility. The institution applies Good Laboratory Practices rules according to the EU and OECD. The team are available to participate in EU validation programmes.

Sources of financing: Proyectos de colaboración universidad/empresa (Fundación Bosch-Gimpera), Programa Nacional de Tecnología de Procesos Quimicos CICY 

Type of Institution: University

Staff:

Ana Martín González, Email: jgf00004@teleline.es, Dr Microbiología, Profesor Titular de Universidad, 15 % time devoted to altern. meth.
Juan Carlos Gutiérrez Fernández, Email: jgf00004@teleline.es, Dr en Microbiología, Profesor Titular de Universidad, 15 % time devoted to altern. meth.
Laura Benítez Rico, Email: jgf00004@teleline.es, Dr Microbiología, Ayudante Universidad, 20 % time devoted to altern. meth.
Sergio Callejas Alejano, Email: jgf00004@teleline.es, Ld Biología, Becario UCM predoctoral
Silvia Díaz, Email: jgf00004@teleline.es, Ld Biología, Estudiante Interno .

Total staff involved in alternative methods is 5 people.

Activities / aims: This university department is mainly involved in basic research, non-regulated applied research and method development; it performs cell biology, molecular biology, toxicology (mechanisms of toxicity, basal cytotoxicity, chronic toxicity, ecotoxicity) and monitoring -chemical, biological- testing on a routine basis and physiology and genetics studies at a research level. Toxins, environmental pollutants and wastes are routinely evaluated; pesticides also being occasionally studied.

The main use of alternative methods is for screening.

Model systems: The model systems used are vegetables (microalgae) and in vitro methods: micro-organisms (ciliated protozoa).

The endpoints employed in vitro are morphology, cell viability, cytoskeleton/membranes/enzyme release, cellular proliferation, metabolic activity, nucleic acids and defence systems.

Experimental systems (examples):

1 Tetrahymena thermofila, growth on microplates, by spectrophotometry, toxicity tests of micotoxins, enterotoxins, toxic, replacement test
2 ciliated (Colpoda steinii and other species), growth on microplates, using flourochromes, toxicity of heavy metals, replacement test

Work lines: Tetrahymena thermophila as a biosensor to detect mycotoxins and enterotoxins. Ciliated protozoa as bioindicators of heavy metals in soil and aquatic ecosystems.

Quality assurance / Validation programmes: The team are available to participate in EU validation programmes.

Sources of financing: DGICYT, Projects CAM, Projects EU, Projects UCM

Type of Institution: University

Staff:

Juán Carlos Illera del Portal, Email: vefis07@sis.ucm.es, Dr Veterinaria, Profesor Titular de Universidad

Total staff involved in alternative methods is 1 person.

Activities / aims: This university department is mainly involved in basic research, method development, method validation and alternatives to animals in education; it performs biochemistry, pharmacology, physiology, toxicology (reproductive cytotoxicity) studies and production (animal) on a routine basis and molecular biology, biokinetics and biotransformation, toxicology (mechanisms of toxicity, acute systemic toxicity, ocular cytotoxicity, nephrotoxicity, immunotoxicity and sensitisation, haematoxicity, hepatotoxicity), nutrition and culture methodology studies at a research level. Hormones and food additives are routinely evaluated; vaccines and wastes also being occasionally studied.

The main use of alternative methods is for screening.

Model systems: The model systems used are conventional animal models and in vitro methods (organ culture).

Work lines: Endocrinology. Food Additives. Reproduction. Immunosupression.

Quality assurance / Validation programmes: The institution applies Good Laboratory Practices rules according to the FDA. The team are available to participate in EU validation programmes in relation to toxicology

Staff:

Concepción García Alfonso, Email: bb1gaslm@uco.es, Dra en Veterinaria, Prof Titular Univer., 50 % time devoted to altern. meth.

Total staff involved in alternative methods is about 1 person.

Activities / aims: This university department is mainly involved in basic research, non-regulated applied research and methods development; it performs biochemistry, toxicology (mecanisms of toxicity, ecotoxicity) and monitoring --chemical, biological- studies on a routine basis. Diverse chemical compounds, pesticides and environmental pollutants are routinely evaluated.

The main use of alternatives methods is for screening and complementary studies.

Model systems: The model systems used are conventional animal models (wild mouse), invertebrates (bivalve), in vitro methods -- cell lines culture (Vero monkey kidney cells).

The endpoints employed are in vitro biotransformation systems (glutathion-S-transferase, EROD, GOR) and defence systems (antioxidative enzymes).

Experimental systems (examples):

1 Vero monkey kidney cells, neutral red uptake, MTT reduction, cell proliferation, antioxidative and biotransformation enzymes, biological effects of environmental pollutants wild mouse liver, antioxidative and biotransformation enzymes, biological effects of environmental compounds

Work lines: The development and use of in vitro methods applied to different materials and areas, particularly the investigation of environmental chemicals induced oxidative stress using Vero monkey kidney cells.

Sources of financing: CICyT, Junta de Andalucia, EU

Type of Institution: University

Staff:

Nicolás Olea Serrano, Email: nolea@goliat.ugr.es, Dr Medicina, Catedrático Universidad, duties related to altern. meth.: Representante Español en EU DG XII y DG XI, OECD en disruptores endocrinos.
Maria F. Olea Serrano, Email: folea@platon.ugr.es, Dr Farmacia, Catedrático Universidad
Rosa Pulgar Encinas, Email: rpulgar@goliat.ugr.es, Dra Medicina, Profesor Ayudante Universidad
Ana Rivas Velasco, Email: arivas@goliat.ugr.es, Ld Farmacia, Becaria MEC
Mariana Fernández Cabrera, Ld Ciencias Químicas, Becaria Fundación HUSC

Total staff involved in alternative methods is 12 people.

Activities / aims: This university department is mainly involved in basic research and non-regulated applied research; it performs cell biology, molecular biology, biochemistry, toxicology (endocrine disruption, chronic toxicity), monitoring -chemical, biological- and culture methodology studies on a routine basis. Hormones, diverse chemical compounds, pesticides, environmental pollutants and wastes are routinely evaluated; pharmaceuticals, cosmetics, medical devices and biomaterials also being occasionally studied.

The main use of alternative methods is for screening.

Model systems: The model systems used are in vitro methods: cell lines culture (MCF7).

The endpoints employed in vitro are cell viability, cellular proliferation and metabolic activity (MTT reduction/SRB).

Experimental systems (examples):

1 E-Screen. MCF7 Cells in culture, cell proliferation, assessment of the total xenoestrogen burden, replacement test
2 MCF7 cells, expression of pS2, estrogenicity
3 MCF7 cells, expression of ER and PgR, estrogenicity
4 Uterine test, organ weight, estrogenicity

Work lines: Endocrine disrupting chemicals (EDCs). Identification of chemicals with hormonal activity. Identification of inadvertent exposure pathways to EDCs in humans. Identification of new chemicals with estrogenic/antiestrogenic activity. Assessment of the total xenoestrogen burden in human samples. Breast cancer and xenoestrogens. Cryptorchidism and male reproductive defects and EDCs. Bisphenols as EDCs.

Quality assurance / Validation programmes: A Quality Assurance Unit is operating inside the facility. The institution applies Good Laboratory Practices rules according to the EU, OECD and the FDA. The team have previously been involved in the following alternative method validation programme: collaborative studies of in vitro tests for estrogenicity 1997-98. They are available to participate in EU validation programmes in relation to test validation programmes for estrogenicity and endocrine disruption

Sources of financing: UE PL5-1129/CICYT, FIS 1959-95, JA 140/94; 159/96; 231/97

Fuente / Source: Guillermo Repetto, Ana del Peso, Manuel Salguero, Manuel Repetto (1999) Inventory of the Spanish Institutions and Scientists Involved in Alternatives to the use of Laboratory Animals (Refinement, Reduction or Replacement) Revista de Toxicología 16: 50-127.