Morfološke promjene C2C12 stanica tijekom procesa diferencijacije

undergraduate thesis

• Title (english): Morphological changes in C2C12 cells during the differentiation process
• Author: Neli Kolar
• Mentor: Višnja Gaurina Srček (mentor)
• Granter: University of Zagreb; Faculty of Food Technology and Biotechnology; (Department of Biochemical Engineering); (Laboratory for Cell Technology, Application and Biotransformations); Zagreb
• Defense date and country: 2023-07-14, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

C2C12 stanice potječu iz satelitskih stanica mišjeg bedrenog mišića i dobivene su izolacijom stanica nakon inducirane ozljede koja potiče regeneraciju. Lako se uzgajaju i diferenciraju u kulturi, te se zbog toga često koriste kao modelni sustav za proučavanje rasta i diferencijacije mišića. Cilj ovog rada bio je praćenje rasta i usporedba morfoloških promjena C2C12 stanica u medijima za proliferaciju i diferencijaciju. Tijekom eksperimenta, nakon što su stanice dosegle konfluenciju od 60 - 70%, dio stanica prebačen je u medij za diferencijaciju koji je sadržavao 2% konjskog seruma, a dio stanica je nastavio rasti u mediju za uzgoj s 10% fetalnog goveđeg seruma. Metodom tripan plavo praćena je dinamika rasta C2C12 stanica tijekom 264 sati, a morfološke promjene su praćene bojanjem stanica bojom kristal-ljubičasto koje su vizualizirane pod inverznim mikroskopom. Na temelju krivulja rasta C2C12 stanica uočeno je da stanice koje su rasle u 2% konjskog seruma ostvaruju 2,5x manji prinos u odnosu na stanice koje su rasle u mediju za proliferaciju s 10% fetalnog goveđeg seruma. Morfološke karakteristike C2C12 stanica u mediju za diferencijaciju pokazuju da je došlo do diferenciranja stanica i nastajanja višejezgrenih, izduženih miotuba. Za to vrijeme, C2C12 stanice koje su rasle u mediju za proliferaciju zadržale su nediferenciranu, mioblastnu morfologiju.

Abstract (english)

C2C12 cells were originally derived from satellite cells from the thigh muscle of female mouse donor after a crush injury. They are easily cultured and differentiated in cell culture and often are used as a model system for studying muscle growth and differentiation. The aim of this thesis was to study the growth and morphological characteristic of C2C12 cells in proliferation and differentiation media. During the experiment, once the cells reached 60-70 % confluence, a part of cells were transferred to a differentiation medium containing 2% horse serum, while the other part remained to grow in culture medium with 10% foetal bovine serum. The growth characteristics of C2C12 cells were determined by Trypan blue method during 264 h, while morphological changes in C2C12 cells were analysed by inverted light microscope after crystal-violet staining. Obtained results showed that cells cultivated in 2% horse serum achieved a 2.5x lower total cell yield in comparison to the cells cultivated in a proliferation medium supplemented by 10% foetal bovine serum. Additionally, morphological changes in C2C12 cells during the processes of proliferation and differentiation were observed. Cultivation of C2C12 cells in a differentiation medium resulted in cell differentiation and the formation of multinucleated, elongated myotubes. At the same time, cells cultured in a proliferation medium retained their typical undifferentiated, myoblastic morphology.

• Keywords: diferencijacija; C2C12 stanice; miotubi; mioblasti; tripan plavo
• Keywords (english): differentiation; C2C12 cells; myotubes; myoblasts; Trypan blue
• Language: croatian
• URN:NBN: urn:nbn:hr:159:785107
• Study programme: Title: Biotechnology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) inženjer/inženjerka biotehnologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-05-06 08:20:56