The "Cell Culture Centre" Laboratory at Istituto Zooprofilattico Sperimentale, Brescia, Italy, began its activity in 1979 with the aim of setting up, scouting and controlling cell cultures of different origin and tissue/organ derivation. At the beginning, the main interest was for cell cultures used in the veterinary field; later on however, the attention extended to cell lines used in biomedical research. Consequently, the laboratory has become a reference Centre for those who work to different purposes with tissue cultures. In this respect, the lab can supply a variety of cell lines to National and International public and private Organizations. For the time being, the collection consists of 424 established cell lines, 54 hybridomas and 57 primary cell cultures. On the whole, there are 35,280 vials of frozen cells.
Due to the progressive expansion of the activity and the increasing number of cell lines routinely manipulated, the various working procedures have been improved and standardized. It is thus important, on the one hand, to protect the staff from the possible risks deriving from cell lines infected with different pathogens and, on the other hand, to prepare and expand, under highly controlled conditions, non-contaminated cell lines. To that goal, a specific department of quality control has been started to determine the characteristics of all the cell lines prepared, amplified and stored.
In addition to preparation, controlled propagation and distribution of the various cell cultures, several tissue culture research projects have been started. Scientific results are regularly published and disclosed in national and international congresses. The technical and scientific expertise shown in different collaborations with public and private medical research organizations is the reason why the "Cell Culture Centre" represents a point of reference for those who work, albeit with different objectives, with tissue cultures.
Cell lines now stored can be schematically subdivided into four different types hereunder reported.
Established cell lines BS CL.......
Tumor cell lines BS TCL......
Hybridomas BS HY.......
Primary cell cultures BS PRC
The cells collected derive from 36 animal species
Along with cell culture preparation and manipulation, a key role is played by the quality controls carried out in order to confirm absence of contaminants and identity of each cell line.
To this purpose, several tests are carried out in compliance with European Pharmacopoeia:
Bacteria, fungi and yeasts: investigated by isolation procedures in appropriate culture media.
Mycoplasma: preliminary screening by Hoechst staining, PCR, ELISA; furthermore, isolation procedures and use of indicator cell lines (official methods) are carried out on the final batch of each cell line.
Adventitious viruses of animal and human origin. They are detected by PCR and isolation tests in cell cultures. Furthermore, a haemadsorbing test with blood red cells is also performed.
Species of origin: evaluated by isoenzyme analysis carried out with a commercial kit.
Tumorigenicity: this characteristic is evaluated by either an in vitro test (inoculation of cell cultures in soft agar medium) or in vivo procedure, based on inoculation into nude mice (nu/nu genotype).
Bacterial endotoxins: investigated by the "Limulus amebocyte lisate test".
The Cell Culture Centre activity is performed in compliance with Quality Norm UNI CEI EN ISO/IEC 17025
CELL CULTURES DERIVED FROM PATIENTS WITH ALZHEIMER'S DISEASE
In the framework of a research project, fibroblast cells of patients with Alzheimer's disease have been isolated, amplified and stored. This biological material has been collected with the purpose of finding markers for early diagnosis.
Alzheimer's disease is the most common form of dementia in the world. Even if the symptoms of the illness are mainly related to the process of progressive neuro-degeneration which develops in particular areas of the brain, it is well known that the biological mechanisms at the basis of its pathogenesis often show systemic characteristics, which can be studied in non-neuronal, easily available cell models. The data obtained through scientific studies on fibroblasts and, more recently, platelets and lymphocytes from Alzheimer's disease patients and control healthy subjects are of great importance. The selected subjects have undergone a series of neurological, haemo-chemical, neuro-radiological tests to assess their health status; all the somatic illnesses, preceding concussions, alcohol abuse, or drug treatments in course were accurately registered. The presence of seriousness of possible cognitive deficiences have been monitored through standard means (MMSE, ADAS, NOI, etc...). The subjects potentially fit for inclusion in the study have been informed as to the aim and method of the research project and the procedure to comply with. Obtaining an informed consensus from the subjects has been the necessary requirement to be included in the research project. The patch tests were performed with a circular scalpel in the deltoid area, while the blood sample for platelets and lymphocytes isolation was drawn in a glucose-citrate buffer solution; the cells of interest were separated by one hour and frozen directly as pellets in liquid nitrogen. Each cell sample is associated to correspondent plasma and genomic DNA samples. For each subject, anamnestic, clinical, genotype APOE data are available in the research study, from both patients and controls. The cell cultures derived from healthy subjects and from patients with Alzheimer's disease are indicated with the following code : BS ALZ.