Primerjava osteogenega potenciala primarnih celic iz različnih perinatalnih tkiv v pogojih in vitro

Novak, Leja (author), Zupan, Janja (mentor), Trojner Bregar, Andreja (comentor)

Kostno tkivo ima omejeno sposobnost regeneracije, zato se v sodobni regenerativni medicini vse bolj uveljavljajo mezenhimske matične/stromalne celice (MSC), ki zaradi svoje multipotentnosti in imunomodulatornih lastnosti predstavljajo obetavno terapevtsko možnost. Pomemben vir MSC so perinatalna tkiva, kot so posteljica, amnijska membrana in popkovnica, saj so lahko dostopna, njihova pridobitev pa ni etično sporna. Kljub številnim raziskavam ostaja vprašanje, ali različni perinatalni viri zagotavljajo primerljiv osteogeni potencial. Namen magistrske naloge je bil primerjati osteogeni potencial 11 vzorcev primarnih MSC, izoliranih iz različnih perinatalnih tkiv, v pogojih in vitro. Celice iz amnijske membrane (5 vzorcev) in posteljice (6 vzorcev) smo izpostavili osteogenemu diferenciacijskemu mediju, medtem ko smo vzorce popkovnice zaradi nezadostnega števila celic izključili iz nadaljnje analize. Osteogeno diferenciacijo smo inducirali z dodatkom deksametazona, askorbinske kisline in β–glicerofosfata ter celice po 21 dneh ovrednotili s histokemijskim barvanjem z alizarin rdeče S in spektrofotometrično kvantifikacijo mineralizacije. Rezultati so pokazali, da osteogena indukcija statistično značilno poveča mineralizacijo zunajceličnega matriksa v primerjavi s kontrolnimi vzorci. Primerjava med različnimi tkivnimi viri je pokazala določeno variabilnost v stopnji mineralizacije, vendar razlike niso bile izrazite, kar kaže na funkcionalno podobnost MSC iz amnijske membrane in posteljice. Analiza uspešnosti diferenciacije je dodatno potrdila, da večina vzorcev doseže kriterij uspešne osteogene diferenciacije, ne glede na tkivni izvor celic. Na podlagi teh ugotovitev lahko sklepamo, da primarne MSC iz amnijske membrane in posteljice v pogojih in vitro uspešno odgovorijo na osteogeno stimulacijo s tvorbo mineraliziranega matriksa. Odsotnost statistično značilnih razlik v stopnji mineralizacije nakazuje na podoben funkcionalen odziv celic iz različnih perinatalnih virov, kar utemeljuje njihovo nadaljnjo raziskovanje v regenerativni medicini.

• Language: Slovenian
• Keywords: mezenhimske matične celice, perinatalna tkiva, osteogeneza, alizarin rdeče S, regenerativna medicina
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: FFA - Faculty of Pharmacy
• Publisher: [L. Novak]
• Year: 2026
• PID: 20.500.12556/RUL-183667
• UDC: 576.7:611.018
• COBISS.SI-ID: 282227203
• Publication date in RUL: 17.06.2026

Secondary language

• Language: English
• Title: In vitro comparison of the osteogenic potential of primary mesenchymal stromal cells derived from different perinatal tissues

Abstract

Bone tissue has a limited regenerative capacity; therefore, mesenchymal stem/stromal cells (MSCs) are becoming increasingly prominent in modern regenerative medicine, representing a promising therapeutic option due to their immunomodulatory properties. Perinatal tissues, such as placenta, amniotic membrane and umbilical cord, are significant sources of MSCs as they are easily accessible and their acquisition is ethically non-controversial. Despite extensive research, the question remains whether different perinatal sources provide comparable osteogenic potential. The aim of this master’s thesis was to compare the in vitro osteogenic potential of 11 primary MSC samples isolated from various perinatal tissues. Cells from the amniotic membrane (5 samples) and placenta (6 samples) were exposed to an osteogenic differentiation medium, while umbilical cord samples were excluded from further analysis due to insufficient cell concentration. Osteogenic differentiation was induced by adding dexamethasone, ascorbic acid and β-glycerophosphate. After 21 days, differentiation was evaluated using alizarin red S histochemical staining and spectrophotometric quantification of mineralization. The results showed that osteogenic induction significantly increases the mineralization of the extracellular matrix compared to control samples. Comparison between the different tissue sources revealed some variability in the degree of mineralization; however, the differences were not pronounced, suggesting functional similarity between MSCs derived from the amniotic membrane and the placenta. Success rate analysis of the differentiation further confirmed that majority of samples met the criteria for successful osteogenic differentiation, regardless of the cell source. Based on these findings, we can conclude that MSCs from amniotic membrane and placenta successfully respond to osteogenic stimulation in vitro by forming a mineralized matrix. The absence of statistically significant differences in the degree of mineralization suggests a similar functional response of cells from different perinatal sources, warranting their further investigation in regenerative medicine.

• Keywords: mesenchymal stem cells, perinatal tissues, osteogenesis, alizarin red S, regenerative medicine