Isolation and characterisation of bovine muscle stem cell progenitors

master's thesis

• Title (croatian): Izolacija i karakterizacija matičnih stanica prekursora mišićnih stanica iz goveda
• Author: Josip Čačković
• Mentor: Petra Kluger (mentor)
• Committee member: Marko Močibob (predsjednik povjerenstva)
• Committee member: Ivan Kodrin (član povjerenstva)
• Committee member: Iva Juranović Cindrić (član povjerenstva)
• Committee member: Jasmina Rokov Plavec (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Chemistry); Zagreb
• Defense date and country: 2023-04-03, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Chemistry

Abstract

This thesis investigates the technical feasibility of bovine muscle progenitor stem cells as a
source for in vitro meat cultivation. The cells were isolated using enzymatic digestion. Bovine
muscle progenitor stem cells were identified using the paired box protein, Pax7 using
immunocytochemical assays and fluorescence microscopy. The cells were treated with
SB203580, a common inhibitor for the p38 mitogen activated protein kinase pathway to
examine its influence on the cell’s proliferation and differentiation capabilities. However, it
didn’t yield the expected outcome. Collagen type 1, gelatine and laminin were used as coatings
and compared to see their influence on the selectivity of the target cell type, of which laminin
was the most promising. Lastly, cells were sorted via fluorescence activated cell sorting using
CD29, CD56 as positive, and CD31 and CD45 as negative cell surface markers, which proved
to be an auspicious method for the purification of the cell culture.

Abstract (croatian)

Ova disertacija istražuje tehničku izvedivost korištenja matičnih stanica prekursora mišićnih
stanica iz goveda za uzgoj mesa in vitro. Stanice su izolirane primjenom enzimatske digestije.
Matične stanice prekursori mišićnih stanica iz goveda identificirane su pomoću transkripcijskog
faktora Pax7, uporabom imunocitokemijskih testova i fluorescentne mikroskopije. Stanice su
tretirane SB203580, inhibitorom metaboličkog puta p38 mitogen-aktivirane proteinske kinaze,
kako bi se ispitao njezin utjecaj na sposobnosti proliferacije i diferencijacije stanica. Međutim,
korištenje istog nije dalo očekivani rezultat. Stanično posuđe tretirano je raznovrsnim
premazima: kolagen tip 1, želatina i laminin, kako bi se promatrao njihov utjecaj na proliferaciju
i selektivnost prema stanicama od interesa. Laminin se pokazao kao najobećavajući. Konačno,
stanice su sortirane putem fluorescentno aktiviranog sortiranja stanica primjenom CD29, CD56
kao pozitivnih i CD31 i CD45 kao negativnih površinskih staničnih markera, što se pokazalo
izvrsnom metodom za pročišćavanje kulture stanica.

• Keywords: cultured meat; characterization; fluorescent activated cell sorting; p38 MAPK; satellite cells; surface coating
• Keywords (english): fluorescentno aktivirano sortiranje stanica; p38 MAPK; satelitske stanice; premazi staničnog posuđa
• Language: english
• URN:NBN: urn:nbn:hr:217:984210
• Study programme: Title: Graduate university studies in chemistry; specializations in: Research programme; Course: Research programme; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra kemije (magistar/magistra kemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2023-04-04 13:01:59