Cell Cultures
OIE Collaborating Centre Reports
Activities in 2013
Title of Collaborating Centre: / Collaborating Centre for Cell CulturesAddress of Collaborating Centre: / Via Bianchi 9, 25124, Brescia, Italy
Tel.: / +39-0302290248
Fax: / +39-0302290386
e-mail address: /
website: / www.izsler.it
Name of Director of Institute (Responsible Official): / Prof. Stefano Cinotti
DVM, Professor
General Manager of IZSLER
Name (including Title and Position) of Head of the Collaborating Centre (formally OIE Contact Point): / Dr. Maura Ferrari
Biologist
Head of Cell Culture Collaborating Centre
Name (including Title and Position) of writer of this report
(if different from above)
Summary of activities specifically related to the mandate of
OIE Collaborating Centres
ToR: To provide services to the OIE, in particular within the region, in the designated specialty, in support of the implementation of OIE policies and, where required, seek for collaboration with OIE Reference Laboratories
ToR: To identify and maintain existing expertise, in particular within its region
1. Activities as a centre of research, expertise, standardisation and dissemination of techniques within the remit of the mandate given by the OIE
Disease control
Epidemiology, surveillance, risk assessment, modelling
Training, capacity building
Zoonoses
Wildlife
Avian diseases
Aquatic animal diseases
Animal welfare
Diagnosis, biotechnology and laboratory
Veterinary medicinal products
Vaccines
Food safety
Feed safety
Other (Name the category: Cell cultures and their application for diagnosis, research, therapy, production and new in vitro tests)
DIAGNOSIS, BIOTECHNOLOGY AND LABORATORYTitle of activity / Scope
Basic activity : preparation, amplification, control, use and distribution of the different types of cell cultures obtained from different sources / Maintain and increase the cell culture types to be used in the different fields of Scientific Community for diagnosis, reasearch and production of viruses from different species. Preparaton of reference standards for controls of cells used in production of biologicals, including vaccines. In 2013 the collection has resulted to be composed by the following cell types:
normal established cell lines No. 224
tumour established cell lines No. 247
hybridomas No. 61
primary cell cultures No. 65
animal Mesenchymal Stromal Cells No. 26
Total No. of :
New Organizations / Cell types: total number 623
Frozen cell vials: number 49,642
Cells that have been distributed: number 336
New Organizations that have requested cells to the OIE Collaborating Centre: number 13 of which 8 national and 5 international (Singapore, United Kingdom, France, Austria, Philippines);
In total 809 are the organizations that over the years have requested cells to OIE Collaborating Centre. 675 of them are national; 121 are of other countries
Other:
Title of activity / Scope
Detection of extraneous agents by cell culture isolation.
Validation of production steps.
Quality controls
Preparation and amplification of new cell cultures
Research activity / Testing of pharmaceutical products, viral vaccines, cells used for vaccine production, mesenchymal stromal cells used for therapy, cells used in virology laboratories in order to ensure their purity.
Evaluation of the efficacy of cleaning, disinfection and sterilization procedures to inhibt virus replication. These tests are carried out according to international guidelines
To ensure the characteristics, identity and purity of the the cell cultures and based on testing methods to evaluate microbiological , mycoplasma and virus contamination. Oncogenic features are also evaluated to ensure the safety of cells used for production of vaccines and biologicals. All testing methods are in accordance to European Pharmacopeia and performed according to quality management system (ISO IEC 17025)
To increase the cell culture collection with the aim to offer a wider possibility of use (isolation of novel virus, development of new serological tests, employment in new fields etc)
It was based on the studies made according to reasearch projects approved with three main different strategies:
1) to improve knowlwdges on the therapeutic application of Mesenchymal Stromal Cells in animals;
2) to develop new testing methods for virus detection and for evaluation of other quality parameters;
3) to develop new in vitro tests to be used as alternative to in vivo experiments conducted on animals.
ToR: To propose or develop methods and procedures that facilitate harmonisation of international standards and guidelines applicable to the designated specialty
2. Proposal or development of any procedure that will facilitate harmonisation of international regulations applicable to the surveillance and control of animal diseases, food safety or animal welfare
Proposal title / Scope/Content / Applicable area /Development of new testing methods
Reference number of the test:
02/308
02/309
02/310
02/311
02/328
02/329
02/330
02/364
02/365
04/099
04/117
05/26
09/171
09/172
09/173
09/174
09/175
09/177
09/189
09/190
09/191
Standardization of the procedures regarding all steps involved in cell culture field / To ensure the purity, safety, and authentity of cell cultures, avoiding to obtain not corrct results regarding testing methods / production / research carried out with cells of no proper quality parameters.
Testing methods developed in 2013:
Evaluation of cell cultures origin by isoenzyme analysis (Authentikit™ system, Innovative Chemistry)
Evaluation of the growth efficiency of animal serum
Evaluation of the transformed features of cell cultures by in vitro test
Tumorigenicity of cell cultures by in vivo test
Evaluation of mesenchymal stromal cells differentiation capacity toward adipocites
Evaluation of mesenchymal stromal cells differentiation capacity toward chondrocytes
Evaluation of mesenchymal stromal cells differentiation capacity towards osteoblasts
Evaluation of bacterial endotoxin levels in biological and pharmaceutical products
Biological evaluation of medical devices by in vitro cytotoxicity assay (MTT)
Detection of anti-PRRSV antibodies by serum neutralization
Detection of adventitious viruses in biological/pharmaceutical products in cell culture
Detection of human cytomegalovirus (HCMV) in biological/pharmaceutical products by PCR real-time (kit “Artus® CMV RG PCR”)
Detection of Epstein-Barr virus (EBV) in biological/pharmaceutical products by PCR real-time (KIT “Artus® EBV RG PCR”)
Detection of Hepatitis B virus (HBV) in biological/pharmaceutical products by PCR real-time (KIT “Artus® HBV RG PCR”)
Detection of Hepatitis C virus (HCV) in biological/pharmaceutical products by RT-PCR real-time (KIT “Artus® HCV RG RT-PCR”)
Detection of herpes simplex virus (HSV) type 1 and type 2 in biological/pharmaceutical products by PCR real-time (KIT “Artus® HSV-1/2 RG PCR”)
Detection of human immunodeficiency virus (HIV) in cell culture and reagents by RT-PCR real-time (KIT “Artus® HI Virus-1 RG RT-PCR”)
Detection of human chondrocytes molecular expression markers (link protein and aggrecan) in pharmaceutical products by PCR
Detection of Swine Classical fever (SCF) in biological and pharmaceutical products by PCR Real-Time
Detection of swine parvovirus (PPV) in biological and pharmaceutical products by PCR Real-Time
The flow chart of the cell culture preparation, cultivation, control and storage is approved by the quality management system and it allows to trace each step of the process. The final product must have high quality parameters and, if possible, its range of application must be evaluated (for instance virus isolation, serological tests, in vitro alternative methods, etc) / Surveillance and control of animal diseases
Food safety
Animal welfare
Other
ToR: To establish and maintain a network with other OIE Collaborating Centres designated for the same specialty, and should the need arise, with Collaborating Centres in other disciplines
ToR: To carry out and/or coordinate scientific and technical studies in collaboration with other centres, laboratories or organisations
3. Did your Collaborating Centre maintain a network with other OIE Collaborating Centres (CC), Reference Laboratories (RL), or organisations designated for the same specialty, to coordinate scientific and technical studies?
Yes No
Name of OIE CC/RL/other organisation(s) / Location / Region of networking Centre / Purpose /Centre de Recerca en Sanitad Animal, CReSa, campus UAB / Barcelona (Spain) / Africa
Americas
Asia and Pacific
Europe
Middle East / to identify cell lines able to allow Porcine reproductive and Respiratory Syndrome Virus – PRRSV isolation from target tissues of infected animals
OIE Reference Laboratory for Avian influenza and Newcastle disease / Padova, Italy / Africa
Americas
Asia and Pacific
Europe
Middle East / to develope of expertise and standardization of the cell culture for use in virology field; use of cell lines as alternative to chichen embryos for virus isolation and amplification
Animal Health and Veterinary Laboratories Agency (AHVLA) / Surrey, UK / Africa
Americas
Asia and Pacific
Europe
Middle East / to develope of expertise and standardization of the cell culture for use in virology field; use of cell lines as alternative to chichen embryos for virus isolation and amplification
OIE Reference Laboratory for swine influenza / IZSLER Parma, Italy / Africa
Americas
Asia and Pacific
Europe
Middle East / to develope of expertise and standardization of the cell culture for use in virology field; use of cell lines as alternative to chichen embryos for virus isolation and amplification
4. Did your Collaborating Centre maintain a network with other OIE Collaborating Centres, Reference laboratories, or organisations in other disciplines, to coordinate scientific and technical studies?
Yes No
Name of OIE CC/RL/other organisation(s) / Location / Region of networking Centre / Purpose /The National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) / London
United Kingdom / Africa
Americas
Asia and Pacific
Europe
Middle East / Coooperation in the translation in italian language of the ARRIVE guidelines used to improve the procedures of in vivo experiments; their disclosure to Organizations involved in this fields
National Reference Centre for Classical and African swine fever / IZS Perugia, Italy / Africa
Americas
Asia and Pacific
Europe
Middle East / Cooperation in suggesting cell lines to be used for virus isolation and cultivation
Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise: Reference Centre for bluetongue virus / IZS Teramo, Italy / Africa
Americas
Asia and Pacific
Europe
Middle East / Involved for field sera biobanking toward bluetongue
The Roslin Institute, University of Edinburgh, (Prof. Tahar Ait-Ali) / Edinburgh
UK / Africa
Americas
Asia and Pacific
Europe
Middle East
National Veterinary Research Institute, Department of Swine Diseases OIE Reference Laboratory for PRRS.
(Dr. Tomasz Stadejek) / Warsaw
Polond / Africa
Americas
Asia and Pacific
Europe
Middle East / Cost Action: viruses related to swine respiratory disease with particular reference to PRRS
University of Nebraska-Lincoln, School of Veterinary and Biomedical Sciences (Prof. Fernando Osorio) / Nebraska / Africa
Americas
Asia and Pacific
Europe
Middle East / Cooperation in pathogenetic studies on PRRRSV virus infection in pigs
ToR: To place expert consultants at the disposal of the OIE.
5. Did your Collaborating Centre place expert consultants at the disposal of the OIE?
Yes No
Name of expert / Kind of consultancy / SubjectDr Maura Ferrari / Author of chapter / Author of Chapter of Molecular Diagnostics in Laboratory Veterinary Practice (in preparation)
Dr. Maura Ferrari / Co-author of chapter / Detection of Neutralizing Antibodies
in Pigs Inoculated with an Inactivated
Vaccine Against Porcine Circovirus Type
2 (PCV2)
ToR: To provide, within the designated specialty, scientific and technical training to personnel from OIE Member Countries
6. Did your Collaborating Centre provide scientific and technical training, within the remit of the mandate given by the OIE, to personnel from OIE Member Countries?
Yes No
If the answer is yes, please provide the total number of trained persons for each of the following categories:
a) Technical visits: 2
b) Seminars:
c) Hands-on training courses:
d) Internships (>1 month):
ToR: To organise and participate in scientific meetings and other activities on behalf of the OIE
7. Did your Collaborating Centre organise or participate in the organisation of scientific meetings on behalf of the OIE?
Yes No
ToR: To collect, process, analyse, publish and disseminate data and information relevant to the designated specialty
8. Publication and dissemination of any information within the remit of the mandate given by the OIE that may be useful to Member Countries of the OIE
(Indicate in the appropriate box the number by category)
a) Articles published in peer-reviewed journals: 6
Renzi S, Riccò S, Dotti S, Sesso L, Grolli S, Cornali M, Carlin S, Patruno M, Cinotti S, and Ferrari M. (2013). Autologous bone marrow mesenchymal stromal cells for regeneration of injured equine ligaments and tendons: a clinical report. Res. Vet. Sci., 95 (1), 272-7.Dotti S, Guadagnini G, Salvini F, Razzuoli E, Ferrari M, Alborali Gl, Amadori M. (2013). Time-course of antibody and cell-mediated immune responses to porcine Reproductive and Respiratory syndrome virus under field conditions. Res Vet Sci., 94, 510-517.
Riccò S, Renzi S, Delbue M, Conti V, Merli E, Ramoni R, Lucarelli E, Gnudi G, Ferrari M, Grolli S. (2013). Allogenic adipose tissue-derived mesenchymal stem cells in combination with platelet rich plasma are safe and effective in the therapy of superficial digital flexor tendonitis in the horse. Int J Immunopathol Pharmacol. 26, 61-68.
Petrini S, Paniccià M, Silenzi V, Ciuti F, Bresaola M, Fortunati M, Perugini G, De Mia GM, perugini G, Ferrari M,, 2013. Detection of neutralizing antibodies in pigs inoculated with an inactivated vaccine against porcine Circovirus type 2 (PCV2). Trends in veterinary sciences: current aspects in veterinary morphophysiology, biochemistry, animal production, food hygiene and clinical sciences. C. Boiti et al. (eds.), Trends in Veterinary Sciences, DOI: 10.1007/978-3-642-36488-4_12, Springer-Verlag, 63-66.
Petrini S, Silenzi V, Manuali E, Paniccià M, Sensi M, Fortunati M, Feliziani F, Ferrari M, De Mia GM, 2013. Association of the main viral pathogens with Porcine Respiratory Disease Complex (PRDC) in Central Italy. Journal of Veterinary Diagnostic Investigation DOI 10.1007/s11259-013-9552-3, Vet Res Commun (2013) 37:251–257
Petrini S, Ramadori G, Villa R, Borghetti P, De Angelis E, Cantoni AM, Corradi A, Amici A, Ferrari M, 2013. Evaluation of Different DNA Vaccines against Porcine Reproductive and Respiratory Syndrome (PRRS) in Pigs. doi:10.3390/vaccines1040463. Vaccines 2013, 1, 463-480.
b) International conference presentations: 3
1) Time-course of antibody and cell-mediated immune responses to porcine reproductive and respiratory syndrome virus under field conditions. 10th International Veterinary Immunology Symposium (IVIS) : 28th August -1 St September 2013 Milan, ItalyAn Italian Biobanking of Veterinary Resources at IZSLER. 7TH Epizone Annual Meeting : 1 - 4 October 2013, Bruxel, Belgium
2) Biobanking of animal Mesenchymal Stromal Cells and quality controls for a safe therapeutic application.European Society Biopreservation and Biobanking (ESBB )Annual Meeting. 8 - 11 October 2013, Verona, Italy
3) Mesenchymal stromal cells (MSCs) alteration following in vitro amplification. Advances in In-Vitro Cell and Tissue Culture and 5th Annual Quasi-Vivo® User Group Meeting. 21 - 22 May 2013, Liverpool, United Kingdom
c) National conference presentations: 5