Primerjava sposobnosti pomnoževanja primarnih celic iz perinatalnih tkiv v pogojih in vitro na dveh različnih gojiščih

Primožič, Andreja (avtor), Zupan, Janja (mentor), Trojner Bregar, Andreja (komentor)

Perinatalna tkiva, med katera uvrščamo amnijsko membrano, posteljico/placento in popkovnico, predstavljajo dostopen in etično sprejemljiv vir mezenhimskih matičnih/stromalnih celic (MSC), ki imajo velik potencial za uporabo v regenerativni medicini. Ključne prednosti perinatalnih MSC vključujejo visoko proliferativno sposobnost, nizko imunogenost ter možnost neinvazivnega pridobivanja. Na uspešnost izolacije in gojenja teh celic v pogojih in vitro vplivajo številni dejavniki, med drugim izvor tkiva, čas od odvzema do obdelave, uporabljena metoda izolacije ter sestava gojišča. Tekom izdelave magistrske naloge je bil naš namen primerjati vpliv dveh različnih gojišč, standardnega LG-DMEM in komercialnega Mesencult®, na uspešnost izolacije ter proliferativno sposobnost MSC iz perinatalnih tkiv. V raziskavo smo vključili vzorce devetih porodnic, pri čemer je vsaka darovala tri vrste tkiv (amnijsko membrano, posteljico in popkovnico). Vzorce smo vzporedno gojili v obeh gojiščih in spremljali čas do pritrditve celic, število tripsiniziranih celic ter sposobnost tvorbe kolonij (CFU-F). Uspešnost izolacije MSC smo določili na podlagi prisotnosti pritrjenih celic, ki so bile sposobne tvoriti kolonije. Rezultati študije kažejo, da je bila izolacija uspešna pri manj kot polovici vseh vzorcev, pri čemer med uporabljenima gojiščema nismo zaznali statistično značilnih razlik. Tudi pri primerjavi števila tripsiniziranih celic in sposobnosti tvorbe kolonij med LG-DMEM in Mesencult® nismo ugotovili statistično značilnih odstopanj, čeprav so bili rezultati med posameznimi vzorci precej variabilni. Dobljeni rezultati kažejo, da izbira gojišča v naši raziskavi ni bistveno vplivala na uspešnost izolacije ali proliferativno aktivnost MSC iz perinatalnih tkiv. Opazili pa smo veliko biološko variabilnost med vzorci, kar nakazuje na pomemben vpliv drugih dejavnikov na rast primarnih celic, kot je na primer izvor tkiva. Za nadaljnje raziskave bi bilo smiselno vključiti večje število vzorcev, analizirati kasnejše pasaže celic ter dodatno ovrednotiti njihove funkcionalne lastnosti.

• Jezik: Slovenski jezik
• Ključne besede: primarne mezenhimske matične/stromalne celice, perinatalna tkiva, gojišče, LG-DMEM, Mesencult®.
• Vrsta gradiva: Magistrsko delo/naloga
• Tipologija: 2.09 - Magistrsko delo
• Organizacija: FFA - Fakulteta za farmacijo
• Založnik: [A. Primožič]
• Leto izida: 2026
• PID: 20.500.12556/RUL-183595
• UDK: 602.9:618.2/.5(043.2)
• COBISS.SI-ID: 281745923
• Datum objave v RUL: 16.06.2026

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Comparison of two different culture media on in vitro expansion of primary cells from perinatal tissues

Izvleček

Perinatal tissues, including the amniotic membrane, placenta, and umbilical cord, represent an accessible and ethically acceptable source of mesenchymal stem/stromal cells (MSCs) with significant potential for use in regenerative medicine. The key advantages of perinatal MSCs include high proliferative capacity, low immunogenicity, and non-invasive collection. The success of isolation and in vitro cultivation of these cells is influenced by several factors, including tissue origin, the time between tissue collection and isolation, the isolation method used, and the composition of the culture medium. The purpose of our thesis was to compare two different culture media, standard LG-DMEM and commercial Mesencult®, on the isolation efficiency and proliferation of MSCs derived from perinatal tissues. We included samples from nine donors, each donated three types of perinatal tissues (amniotic membrane, placenta, and umbilical cord). The samples were cultured in parallel in both media, and parameters such as time to cell attachment, number of trypsinized cells, and colony-forming ability (CFU-F) were evaluated. The success of MSC isolation was defined by the presence of adherent cells that were capable to form colonies. Our results showed that isolation was successful in less than half of the samples, with no statistically significant differences detected between the two culture media. Similarly, no statistically significant differences were found when comparing the number of trypsinized cells or colony-forming ability between LG-DMEM and Mesencult®, although the results were variable between individual samples. The results also indicate that, under the conditions of this study, the choice of culture medium did not significantly influence the isolation efficiency or proliferative activity of MSCs derived from perinatal tissues. Moreover, we observed high biological variability between samples, which suggest a significant impact of other factors, such as tissue origin. Future studies should include a larger number of samples, analysis of later cell passages, and further evaluation of functional properties of the cells.

• Ključne besede: primary mesenchymal stem/stromal cells, perinatal tissues, culture medium, LG-DMEM, Mesencult®.