Naslov
Tissue and cell biobank of patients with inborn errors of metabolism in Croatia

Autori
Zekušić, Marija ; Fumić, Ksenija

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Future Medicine (Regenerative Medicine) 2015 / - , 2015, 96-296

Skup
World Conference on Regenerative Medicine, 2015

Mjesto i datum
Leipzig, Njemačka, 21.10.2015. - 23.10.2015

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
skeletal myoblast; skin fibroblast; biobanking

Sažetak
Objectives Rare diseases are a clinically heterogeneous group of disorders with prevalence less than 5/10000. Only 6-8% of the world’s population is likely to be affected by some form of rare disease. Since these diseases affect multiple organ systems and do not have characteristic symptoms, interdisciplinary cooperation is essential to establish diagnosis and include appropriate treatment. Life expectancy in these patients is often reduced ; however, if detected in time, many of these diseases can be successfully treated. Inherited metabolic diseases form a small group of rare diseases diagnosed not only using conventional biological material (blood, urine, cerebrospinal fluid) but also tissue biopsy specimens are often included in the diagnostic process. In the Clinical Unit for Hereditary Metabolic Diseases of the University Hospital Centre Zagreb, different types of progenitor cells (dermal fibroblasts and skeletal myoblasts) and various human biopsy specimens (muscle, liver) of patients with rare hereditary metabolic diseases have been collected, processed, stored and handled for many years. Until now, Croatia had no organized biobank that would meet common European standards and is currently in process of establishing the first non-profit tissue and cell biobank of patients with inborn errors of metabolism. The main goal is to optimize the establishment of cell cultures and ultimately implement and harmonize Standard Operating Procedures in accordance with the European Network of Biobanks (EuroBioBanks, EBB) and ISO 15189 standards. Materials and Methods Progenitor cells were isolated from skin and skeletal muscle biopsy specimens of patients with inborn errors of metabolism. After the establishment of primary and secondary cultures, the cells were frozen in liquid nitrogen. Dulbecco's modification of Eagle's medium supplemented with 20% of fetal bovine serum (FBS), 1% of L-glutamine and 1% of antibiotics was used in the cultivation of fibroblasts. Myoblasts, with a small number of fibroblasts, were cultured in a Skeletal Muscle Growth Medium, SGM (PromoCell, Germany) supplemented with 5% of growth factors (epidermal growth factor, fibroblast growth factor and insulin). Into the medium 10% of FBS, 1.5% of glutamine and antibiotics were added. Purification of myoblasts from fibroblasts was performed using Magnetic- Activated Cell Sorting Technology (Miltenyi Biotec, Germany). IgG-microbeads coated in specific primary antibody NCAM/CD56 were used to label myoblasts and separate them from unlabelled cells. Purity of myoblasts obtained by immunomagnetic sorting was assessed using flow cytometer before the storage in liquid nitrogen. Results The number of established cell cultures (1002 human fibroblast cultures and 48 skeletal myoblast cultures) together with the number of human biopsy specimens (337) isolated from patients with rare hereditary diseases was entered in the first Croatian biobank database. The purity of myoblast cell cultures was nearly 90%. The entire process of collecting, processing, storage and handling of samples was performed in concordance with valid European standards. Conclusion The first step in establishment of Croatian tissue and cell biobank has been taken. Requirements for data networking and harmonization of Croatian biobank according to the EBB and ISO 15189 standards have been accomplished. Cell culture of human myoblasts opens up new possibilities in the field of research of adult stem cells.

Izvorni jezik
Engleski

Znanstvena područja
Kliničke medicinske znanosti

POVEZANOST RADA