Telethon ha contribuito con un finanziamento alla creazione di una banca di "Mioblasti provenineti da pazienti affetti da malattie neuromuscolari " (Progetto n° C. 56).
Il progetto è portato avanti in collaborazione con il Dott. Lorenzo Maggi della Divisione delle Malattie Neuromuscolari, presso l'Istituto Neurologico "Carlo Besta", Via Celoria 11, 20133 Milano.
Per Contattarci: Tel 02-2394413, 02-2394632 - FAX 02-70633874
Organizzazione della Banca Telethon
Myoblasts are an important renewable source of patient material for protein, DNA and RNA studies, both for diagnostic purposes and for use in various fields of basic and pre-clinical research.
Furthermore, as new genetic tests on blood samples will become available, performing muscle biopsies for diagnostic purposes will be less and less ethic. Therefore, established muscle cell cultures will represent an important and, perhaps, the only source of material for studies in the field muscle diseases.
The bank was started in 1993 both as requirement and result of the routine activity of the Division of Neuromuscular Diseases. Most specimens derive from this activity; a few are sent to the laboratory from other Institutions.
We perform about 8-10 muscle biopsies per week. These are obtained after written informed consent of the patients or the parents. It is emphasized that consent is given to store samples for future research use, and for new diagnostic tests whenever they become available; that the identity of the patient remains confidential; that the material will always be available to the patient; and that the material may be lost due to storage failure.
For most biopsies two samples are frozen in isopentane/liquid nitrogen for histology and biochemistry. One small fragment is fixed for electron microscopy. Another small fragment is collected in sterile medium for myoblast culture; this fragment can either be processed immediately for cell culture, or processed for storage for future cell culture.
For each patient undergoing muscle biopsy, a blood sample for DNA extraction is taken.
Muscle cells and cryoconserved vital biopsies are stored in 2 ml cryotubes in liquid nitrogen. The cryotubes are labelled with a code number, the passage number (where appropriate) and the date of freezing. The code numbers can be decoded (patient identified) only by members of the staff of the muscle pathology Division. Hence all our samples are identifiable and not identified.
At present we have three containers (capacity 6000 samples), one for biopsy specimens and two for myoblasts.
For each container we keep a register indicating the contents of each box and each rack. The data are also stored in Excel format on a computer; both registers are updated regularly; records are kept under lock and key.
For each muscle biopsy from which myoblasts or cryoconserved muscle specimens have been obtained, we also keep slides of stained cryosections, resin blocks of embedded tissue for electron microscopy studies, and histopathological records. From most patients who have undergone muscle biopsy at our Department, detailed clinical data are stored in a database, and will be available on application after consent of the treating physician. Database security is ensured by use of passwords to access data and computers.
There is no possibility at present for unauthorized third parties to access this information via direct internet links.
All laboratories of the National Neurological Institute C. Besta, including those of the Division of Neuromuscular Diseases, have recently obtained the certification UNI EN ISO 9001/2000. To do so all of them have gone through a series of formalization of procedures, including those concerning security, recording and access of patient data, banking procedures and informed consent.
Cells are checked on a regular basis for bacterial, fungal, mycoplasma and viral contamination.
Care is taken not to cross contaminate cell lines. Cell growth in culture, population doubling times and medium suitability, are regularly monitored by cell counting.
In the past five years, several researchers, either financed by Telethon or by other entities, have required cells from the bank. Usually the researcher who needs them contacts the curator of the bank because he/she knows personally the curator or because the bank is listed as Telethon facility.
The Muscle Cell laboratory of the Division of Neuromuscular Diseases has recently opened a web site with a web page dedicated to the myoblast bank and links to Telethon facilities. Furthermore, the laboratory has become member of a European network concerned with promotion of links between Biological Resource Centres in the field of rare diseases. Both these activities will increase the awareness of the existence of the bank facility among researchers and physicians.
In the past few years an average of 10-15 cell lines per year have been given away to scientists for research and for diagnostic purposes and 2-3 cell lines entered the bank from sources outside the Division. Approximately 50 new cell lines per year have been established from muscle biopsies performed at the Besta Institute.
Publications with acknowlegment
Fibbi G, Barletta E, Dini G, Del Rosso A, Pucci M, Cerletti M, Del Rosso M. Cell invasion is affected by differential expression of the urokinase plasminogen activator/urokinase plasminogen activator receptor system in muscle satellite cells from normal and dystrophic patients. Lab Invest. 2001 Jan;81(1): 27-39.
Fibbi G, D'Alessio S, Pucci M, Cerletti M, Del Rosso M. Growth factor-dependent proliferation and invasion of muscle satellite cells require the cell-associated fibrinolytic system. Biol Chem. 2002 Jan;383(1):127-36.