Diferencijacija mišićnih stanica C2C12 u prisustvu hidrolizata proteina lana

undergraduate thesis

• Title (english): Differentiation of muscle cells C2C12 in presence of flaxseed protein hydrolysate
• Author: Anamarija Šest
• 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: 2024-07-10, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Biljni hidrolizati proteina bogati su izvor peptida i ostalih bioaktivnih spojeva zbog čega se u znanstvenim istraživanjima ispituje njihovo djelovanje na održavanje i rast stanica u kulturi. U ovom eksperimentu ispitano je djelovanje hidrolizata proteina lana na vijabilnost i diferencijaciju mišićnih stanica C2C12. Isto tako ispitano je i posjeduje li hidrolizat proteina lana protektivni učinak na atrofiju C2C12 stanica izazvanu tretmanom deksametazonom. Stanice C2C12 tretirane su hidrolizatom proteina lana u rasponu koncentracija 0,5 – 5 mg/mL i određena je vijabilnost stanica nakon 48 h. U cilju određivanja protektivnog učinka hidrolizata proteina lana, stanice su tretirane s 1 mg/mL hidrolizata te je kroz sedam dana praćen njegov utjecaj na diferencijaciju mišićnih stanica. Protektivni učinak 1 mg/mL hidrolizata na atrofiju C2C12 stanica izazvanu tretmanom 100 μM deksametazona praćen je tijekom dva dana. Metodom bojanja tripan plavo određen je broj stanica, vijabilnost stanica određena je MTS metodom, a učinak na diferencijaciju određen je mjerenjem promjera stanica programom Digycyte i bojanjem stanica bojom kristal – ljubičasto. Na temelju dobivenih rezultata, vidljiv je inhibirajući učinak hidrolizata lana na vijabilnost stanica C2C12 u rasponu od 7,89-17,37 %. Rezultati su pokazali i smanjenje promjera diferenciranih miotuba tretiranih hidrolizatom za 10 %. Usporedbom morfologije kontrolnih i tretiranih stanica s 1 mg/mL hidrolizata lana, uočeno je kako tretirane stanice pokrivaju manju površinu za rast i prisutne su u obliku okruglih nakupina u odnosu na netretirane stanice. Također, uočen je i blagi protektivni učinak hidrolizata lana (+6,5 %) na atrofiju C2C12 stanica u usporedbi sa stanicama tretiranim samo sa 100 μM deksametazona.

Abstract (english)

Plant protein hydrolysate are a rich source of peptides and other bioactive substances which is the reason why scientific research examines their effect on the maintenance and growth of cultured cells. This experiment examines the effect of flaxseed protein hydrolysate on the viability and differentiation of muscle cell C2C12. It was also tested whether the hydrolysate has protective effect on C2C12 atrophy induced by dexamethasone treatment. C2C12 cells were treated with flaxseed protein hydrolysate in concentration range of 0,5 – 5 mg/mL and viability was determined after 48 h. In order to determinate protective effect of flaxseed protein hydrolysate, the cells were treated with 1 mg/mL hydrolysate and its effect on differentiation of muscle cells was monitored over seven days. Protective effect of 1 mg/mL hydrolysate on C2C12 cell atrophy induced by dexamethasone treatment was monitored for two days. The number of cells was determined by Trypan blue method, viability of cells was determined by MTS method, and the effect on differentiation was determined by measuring diameter with program Digicyte and by staining cells with crystal - violet. Based on the obtained results, the inhibiting effect of flaxseed protein hydrolysate on viability of cells is visible in range of 7,89 - 17,37 %. The results also showed a 10 % reduction in the diameter of differentiated myotubes treated with the hydrolysate. By observing the morphology of untreated and treated cells with 1 mg/ml hydrolysate, it was noticed that treated cells cover a smaller area for growth and form a round clusters in relation to untreated cells. In adition, a mild protective effect of flaxseed hydrolysate was observed (+6,5 %) on C2C12 cells atrophy compared to cells treated only with 100 μM dexamethasone.

• Keywords: atrofija; C2C12 stanice; diferencijacija; hidrolizat proteina lana; vijabilnost
• Keywords (english): atrophy; C2C12 cells; differentiation; flaxseed protein hydrolysate; viability
• Language: croatian
• URN:NBN: urn:nbn:hr:159:737087
• 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: 2024-11-11 13:47:31