20.500.12556/RUL-186055 Vpliv zunajceličnega matriksa na diferenciacijo skeletnomišičnih celic v razmerah in vitro Effect of extracellular matrix on differentiation of skeletal muscle cells under conditions in vitro Komponente zunajceličnega matriksa pomembno vplivajo na rast in diferenciacijo skeletnomišičnih celic. V eksperimentih in vitro si prizadevamo doseči zrelejšo obliko mišičnih vlaken, ki kar najbolje odražajo stanje in vivo. Zato se pogosto uporabljajo podlage kot sta Matrigel in Geltrex, ki posnemata sestavo in lastnosti zunajceličnega matriksa. Namen diplomske naloge je bil ugotoviti vpliv Matrigela in Geltrexa na označevalce diferenciacije primarnih človeških skeletnomišičnih celic. Z metodo prenosa western smo določili količine dezmina, MYH7, cepljene kaspaze 3, ter α1, α2 in β1 podenot natrij-kalijeve črpalke (ATP1A1, ATP1A2, ATP1B1). Celice, gojene na podlagah Matrigel in/ali Geltrex, so vsebovale višjo celokupno količino proteinov, višje količine dezmina, MYH7 ter ATP1A1 izooblike natrij-kalijeve črpalke kot celice, gojene neposredno na polistirenu, kar bi lahko pomenilo izboljšano diferenciacijo. Prisotnost cepljene kaspaze 3 ni bila odvisna od tipa podlage, pač pa od gostote celic. Višja gostota celic je bila povezana z višjo količino cepljene kaspaze 3. Z metodo RT qPCR, kjer je endogeno kontrolo predstavljala peptidilprolil izomeraza A, smo določili raven mRNA MYH1, MYH7 in DES. Celokupna količina izolirane RNA je bila višja pri pogojih Matrigel in Geltrex kot pri polistirenu, a statistično značilnih razlik v količini mRNA za dezmin, MYH1 in MYH7 med podlagami ni bilo. Rezultati nakazujejo na pozitiven vpliv podlag Matrigel in Geltrex na diferenciacijo primarnih človeških skeletnomišičnih celic in predstavljajo izhodišče za nadaljne raziskave optimizacije in vitro modelov skeletne mišice. Components of extracellular matrix significantly influence the growth and differentiation of the extracellular matrix. The aim of experiments in vitro is to obtain a mature form of muscle fibers which most closely resemble the state in vivo. Therefore, substrates such as Matrigel and Geltrex, which mimic the composition and properties of the extracellular matrix, are frequently used. Our aim was to compare growth and differentiation markers of primary human skeletal muscle cells in wells without coating and in wells coated with Matrigel or Geltrex. Using Western blot, we determined the abundance of desmin, myosin heavy chain, α1, α2 and β1 subunits of Na+/K+ ATPase (ATP1A1, ATP1A2, ATP1B1) and cleaved caspase 3. The abundance of desmin, MYH7 and α1 subunit of Na+/K+ ATPase were significantly higher in cells grown on Matrigel or Geltrex suggesting enhanced growth and differentiation. Abundance of cleaved caspase 3 seemed to be dependent on the density of cells and not the culturing conditions. Protein concentration measurements showed significantly increased concentration in coated wells. Using RT qPCR with peptidylpropyl isomerase A as an endogenous control, we determined that the mRNA expression levels of MYH7, desmin and Na+/K+ ATPase did not significantly differ between the groups, however, the total mRNA was higher in coated wells. Our results indicate that Matrigel and Geltrex positively influence the differentiation of primary human skeletal muscle cells and provide a basis for further research into the optimization of in vitro models of skeletal muscle. miozin dezmin natrij-kalijeva ATPaza Matrigel Geltrex myosin desmin sodium-potasssium ATPase Matrigel Geltrex true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2026-08-26 10:30:04 2026-08-26 10:30:15 2026-09-09 08:11:22 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 25677 290436611 Kociper_Debora_-_Vpliv_zunajcelicnega_matriksa_na_diferenciacijo_skeletnomisicnih_celic_v_razmer.pdf Kociper_Debora_-_Vpliv_zunajcelicnega_matriksa_na_diferenciacijo_skeletnomisicnih_celic_v_razmer.pdf 1 54FADA81CD33853C49FCA6DCC736AD0B 9a910e65156cd841b37ae1d24c8f5d33a0544a3b5f2a8b552427a702d1a4a97a f32177f6-a127-11f1-9b0d-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=245107 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0