Evaluation of mesenchymal stem cells/ stromal cells chondrogenic potential using Bern score in patients with early degenerative changes

Mezenhimske matične celice so nediferencirane celice z veliko sposobnostjo diferenciacije v tkiva mezodermalnega izvora in visokim potencialom za uporabo v imunomodulatorni in regenerativni terapiji, na primer pri osteoartrozi. Tu gre za degenerativno bolezen sklepov, ki se začne z mikroskopskimi spremembami v hrustancu, ki vodijo v njegovo prekomerno obrabo in sčasoma tudi v poškodbo kosti. V tej magistrski nalogi smo nameravali oceniti sposobnost matičnih celic, da tvorijo hrustančno tkivo v pogojih in vitro. Matične celice smo pridobili iz vzorcev tkiv, ki so se med seboj razlikovali glede na patologijo darovalca (z ali brez degenerativnih sprememb kolka) in mesto odvzema (vzorci sinovije na treh predelih kolka ter vzorec trabekularne kosti). Tkivne vzorce smo pridobili od 15 bolnikov, napotenih na Ortopedsko kliniko UKC Ljubljana. Primarne celice smo gojili v pogojih in vitro bodisi v hondrogenem ali kontrolnem mediju, iz tako nastalih hondrogenih pelet pa pripravili obarvane preparate ter jih nato pregledali pod mikroskopom. Morfologijo pelet smo ocenili z uporabo metodologije po Bernu, izmerili pa smo tudi njihov premer. S statistično analizo smo dokazali vpliv sestave gojitvenega medija na morfologijo pelet ter vpliv mesta odvzema tkiv na premer pelet, medtem ko patologija kolka ni vplivala na nobenega izmed preiskovanih parametrov. Domnevamo, da so ključen vzrok za dobljene rezultate specifične lastnosti mezenhimskih matičnih celic in njihovega mikrookolja ter relativno nizko število vzorcev. Zato verjamemo, da bo za kakovostno oceno sposobnosti hondrogeneze mezenhimskih matičnih celic potrebna izvedba nadaljnjih sorodnih študij z večjim številom vzorcev ter v pogojih z večjo podobnostjo stanju in vivo.Mesenchymal stem cells are non-differentiated cells with a high capacity to differentiate into tissues of mesodermal origin and a high potential for use in immunomodulatory and regenerative therapy, e.g. osteoarthritis. This is a degenerative disease of the joints that begins with microscopic changes in cartilage that lead to its excessive wear and eventually to bone damage. In this work, we aimed to investigate the ability of mesenchymal stem cells to form cartilage under in vitro conditions. We obtained stem cells from samples that differed by the underlying pathology of the donor (with or without degenerative changes of the hip joint) and by the site of collection (sinovium samples from three sites in the hip joint and samples from trabecular bone tissue). Samples were collected from 15 patients at the Orthopaedic Clinic of the University Medical Centre Ljubljana. Primary cells were cultured under in vitro conditions in either hondrogenic or control medium, and the hondrogenic pellets formed were used to prepare stained slides, which were then examined under the microscope. The morphology of the pellets was evaluated using the Bern score, while their diameter was also measured. Using statistical analysis, we demonstrated the influence of growth medium on pellet morphology and the influence of collection site on pellet diameter, while pathology had no effect on either parameter. We hypothesise that the key to these results lies in the specific characteristics of the mesenchymal stem cells and their microenvironment, and in the relatively small number of samples. Therefore, we believe that a larger number of samples, as well as conditions more similar to those in vivo, are needed for a better evaluation of the cartilage-forming ability of mesenchymal stem cells