Vrednotenje učinkovitosti krioprezervacije primarnih celic iz kosti golenice kadavrov

Čuturić, Lidija (author), Zupan, Janja (mentor), Haring, Gregor (comentor)

Krioprezervacija omogoča dolgotrajno shranjevanje celic, vendar lahko vpliva na njihove funkcionalne lastnosti. Namen magistrske naloge je bil ovrednotiti večletno krioprezervacijo primarnih človeških celic iz kostnega tkiva, primerjati sposobnost tvorbe kolonij med sveže izoliranimi in krioprezerviranimi celicami ter preveriti njeno povezanost s trajanjem krioprezervacije. V raziskavo je bilo vključenih 23 vzorcev primarnih celic iz kadavrskega kostnega tkiva, ki je vključevalo mineralizirano trabekularno kost in kostni mozeg. Sveže izolirane celice so bile analizirane v dveh replikatih. En replikat je bil takoj po izolaciji gojen in analiziran v pogojih in vitro. Celice iz drugega replikata so bile po izolaciji neposredno krioprezervirane brez predhodnega gojenja, zato so bile po odmrznitvi obravnavane kot pasaža nič. Shranjene so bile približno od 4,2 do 7,5 let. Po odmrznitvi smo spremljali njihovo sposobnost pritrditve na podlago, morfologijo, proliferacijo in tvorbo kolonij ter rezultate primerjali s pripadajočimi sveže izoliranimi vzorci. Po odmrznitvi so celice tvorile kolonije pri 13 od 23 analiziranih vzorcev, kar predstavlja 56,5 % uspešno izoliranih vzorcev. To kaže, da so nekatere celice tudi po več letih shranjevanja ohranile sposobnost plastične adherence, proliferacije in tvorbe kolonij. Analiza 12 parov sveže izoliranih in krioprezerviranih vzorcev ni pokazala statistično značilnih razlik v sposobnosti tvorbe kolonij (p = 0,1763). Kljub temu je bila med skupinama opazna razlika v porazdelitvi rezultatov, saj je bilo pri krioprezerviranih vzorcih več ničelnih vrednosti. Prav tako ni bila ugotovljena statistično značilna povezava med dolžino krioprezervacije in sposobnostjo tvorbe kolonij (ρ = 0,04127; p = 0,8956). Celice so pri delu vzorcev tudi po večletni krioprezervaciji ohranile sposobnost pritrjanja na podlago, proliferacije in tvorbe kolonij. Statistično značilne razlike med sveže izoliranimi in krioprezerviranimi vzorci nismo dokazali, čeprav je bilo pri slednjih več ničelnih vrednosti. Povezave med trajanjem krioprezervacije in sposobnostjo tvorbe kolonij prav tako nismo dokazali.

• Language: Slovenian
• Keywords: mezenhimske matične/stromalne celice, krioprezervacija, primarne človeške celice, kostni mozeg, sposobnost tvorbe kolonij
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: FFA - Faculty of Pharmacy
• Publisher: [L. Čuturić]
• Year: 2026
• PID: 20.500.12556/RUL-185974
• UDC: 576.3(043.2)
• COBISS.SI-ID: 288920067
• Publication date in RUL: 25.08.2026

Secondary language

• Language: English
• Title: Evaluation of the cryopreservation efficiency of primary cells from cadaveric subchondral bone

Abstract

Cryopreservation enables the long-term storage of cells but may affect their functional properties. The aim of this Master’s thesis was to evaluate the effects of multi-year cryopreservation on primary human cells derived from bone tissue, compare colony-forming capacity between freshly isolated and cryopreserved cells, and assess its association with the duration of cryopreservation. The study included 23 cadaveric samples of primary cells derived from bone tissue, comprising mineralized trabecular bone and bone marrow. Freshly isolated primary cells were analysed in two replicates. The first replicate was culture expanded immediately following cell isolation and analysed. The cells in the second replicate were cryopreserved immediately after isolation without prior culture and were therefore, considered passage zero upon thawing. They were stored for approximately 4.2 to 7.5 years. After thawing, we monitored their propensity for plastic adherence, morphology, proliferation, and colony formation, and compared the results with those of corresponding freshly isolated samples. After thawing, the cells formed colonies in 13 of 23 samples, representing 56.5% of successfully isolated samples. This indicates that some cells retained their ability to adhere, proliferate, and form colonies even after several years of storage. Analysis of 12 pairs of freshly isolated and cryopreserved samples did not reveal statistically significant differences in colony-forming ability (p = 0.1763). Nevertheless, a noticeable difference in the distribution of results was observed between the two groups, as there were more zero values in the cryopreserved samples. Likewise, no statistically significant correlation was found between the duration of cryopreservation and the ability to form colonies (ρ = 0.04127; p = 0.8956). In some samples, cells retained their propensity for plastic adherence and their ability to proliferate and form colonies even after multi-year cryopreservation. No statistically significant difference was demonstrated between freshly isolated and cryopreserved samples, although more zero values were observed in the latter group. No association between the duration of cryopreservation and colony-forming capacity was demonstrated.

• Keywords: mesenchymal stem/stromal cells, cryopreservation, primary human cells, bone marrow, colony-forming ability