Vpliv izbranih dejavnikov na proliferacijo in diferenciacijo mezenhimskih matičnih celic iz maščobnega tkiva psov in mačk

Voga, Metka (author), Majdič, Gregor (mentor), Nemec, Ana (comentor)

Matične celice so v zadnjih letih zaradi svojih edinstvenih lastnosti postale vir razvoja regenerativne medicine in tkivnega inženirstva. Ker različne lastnosti mezenhimskih matičnih celic (MSC) v pogojih in vitro lahko vodijo v potencialno različen terapevtski učinek, je temeljnega pomena odkrivanje vplivov različnih dejavnikov na značilnosti MSC. Uveljavljena podlaga za gojenje MSC je plastična osnova, vendar se zaradi boljšega posnemanja naravnega celičnega okolja za gojenje celic vedno bolj poudarja pomen uporabe celičnih nosilcev. V doktorski disertaciji smo raziskavo sistematično razdelili na dva dela. V prvem delu smo celice gojili na običajni plastični podlagi in proučevali vpliv vrste, spola in poznih pasaž na proliferacijski in diferenciacijski potencial MSC iz maščobnega tkiva (AMSC) psov in mačk. V drugem delu smo celice gojili na celičnih nosilcih iz svilnega fibroina (SF) in proučevali, kako SF vpliva na proliferacijski in diferenciacijski potencial AMSC. Rezultati prvega dela naše raziskave so pokazali, da AMSC mačk izkazujejo slabši proliferacijski in diferenciacijski potencial kot AMSC psov in da tudi pozne pasaže bolj negativno vplivajo na AMSC mačk kot na AMSC psov. Dodatno smo pokazali, da AMSC mačk v manjši meri izražajo preiskovane celične označevalce in v manj optimalnih pogojih izkazujejo slabšo živost. Z rezultati prvega dela raziskave smo pokazali, da vrsta živali lahko bistveno vpliva na značilnosti AMSC v pogojih in vitro in da bi živalsko vrsto morali upoštevati kot spremenljivko pri pripravi celic za regenerativno zdravljenje. Rezultati drugega dela naše raziskave so pokazali, da SF omogoča adhezijo celic in vodi usodo AMSC v smeri hondrogene diferenciacije, ki smo jo potrdili z analizo celične morfologije, barvanjem zunajceličnega matriksa diferenciranih celic ter kvantifikacijo izražanja genov, značilnih za hrustančne celice. Z rezultati drugega dela raziskave smo pokazali, da je hondrogeno diferenciacijo celic moč doseči brez posebnih pogojev gojenja celic, ki so sicer potrebni za hondrogenezo MSC na običajni plastični podlagi. Rezultati doktorske disertacije omogočajo poglobitev pomena poznavanja razlik matičnih celic med živalskimi vrstami in hkrati predstavljajo pomembno osnovo za prihodnje študije in potencial za uvajanje novih zdravljenj z matičnimi celicami v veterinarski klinični medicini.

• Language: Slovenian
• Keywords: mezenhimske matične celice, celična proliferacija, celična diferenciacija, celične kulture, biomateriali, svilni fibroin, hondrogeneza, psi, mačke
• Work type: Doctoral dissertation
• Typology: 2.08 - Doctoral Dissertation
• Organization: VF - Veterinary Faculty
• Place of publishing: Ljubljana
• Year: 2021
• Number of pages: 114 str.
• PID: 20.500.12556/RUL-127692
• Publication date in RUL: 20.06.2021

Secondary language

• Language: English
• Title: Effect of selected factors on proliferation and differentiation of canine and feline adipose derived mesenchymal stem cells

Abstract

Due to their unique properties, stem cells have become an important source for the development of regenerative medicine and tissue engineering in recent years. Since different properties of mesenchymal stem cells (MSCs) in vitro can lead to potentially different therapeutic effects, investigating the influence of various factors on the properties of MSCs is essential. The established basis for the cultivation of MSCs is a plastic surface, but due to the better mimicking of the natural cellular environment, the use of cell carriers for cell cultivation is being increasingly developed. In this dissertation, our studies are systematically divided into two parts. In the first part, cells were grown on a conventional plastic surface and the influence of species, sex and late passages on the proliferation and differentiation potential of canine and feline adipose derived MSCs (AMSCs) were investigated. In the second part, cells were grown on silk fibroin (SF) cell carriers and influence of SF on the AMSCs was investigated. The results of the first part of our study showed that feline AMSCs had poorer proliferation and differentiation potential than canine AMSCs and that late passages had a more negative effect on feline AMSCs than on canine AMSCs. In addition, feline AMSCs expressed fewer examined cell surface markers and exhibited lower viability under less optimal conditions. The results of the first part of the study showed that animal species can significantly affect the properties of AMSCs in vitro and that animal species should be considered when preparing cells for regenerative treatment. The results of the second part of our study showed that SF enables cell adhesion and directs AMSC fate towards chondrogenic differentiation, which was confirmed by the analysis of cell morphology, staining of extracellular matrix of differentiated cells and quantification of genes, characteristic of chondrocytes. With the results of the second part of the study, we have shown that chondrogenic AMSC differentiation can be achieved without the special cell culture conditions that are otherwise required for chondrogenesis of AMSCs on a conventional plastic surface. The results of the dissertation allow us to deepen the knowledge about the importance of the stem cell differences between species and at the same time represent basis for further studies and the potential for the introduction of new stem cell therapies in clinical veterinary medicine.

• Keywords: mesenchymal stem cells, cell proliferation, cell differentiation, cell culture techniques, biomaterials, silk fibroin, chondrogenesis, dogs, cats