Vijabilnost, stanični ciklus i stanična smrt CHO DP-12 stanica tijekom uzgoja uz dodatak hidrolizata lana i konoplje

master's thesis

• Title (english): Cell viability, cell cycle and cell death in CHO DP-12 cells cultivated in media with addition of flax and hemp hydrolysates
• Author: Paula Inkret
• Mentor: Kristina Radošević (mentor)
• Committee member: Andreja Leboš-Pavunc (član povjerenstva)
• 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: 2021-07-22, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Biljni hidrolizati se zadnjih desetljeća istražuju kao potencijalna zamjena za životinjski serum, nepoželjan dodatak mediju za uzgoj proizvodnih staničnih linija. Cilj ovoga rada je istražiti in vitro utjecaj proteinskih hidrolizata lanene i konopljine pogače na rast i vijabilnost CHO DP-12 stanične linije u suspenzijskom uzgoju. Prati se utjecaj 7 hidrolizata različitih koncentracija i frakcija tijekom suspenzijskog uzgoja i mehanizam njihovog citotoksičnog djelovanja. Hidrolizati nisu značajno utjecali na CHO DP-12 staničnu liniju, osim dva ispitana hidrolizata konoplje u koncentraciji od 2 g Lˉ¹. Nefrakcionirani hidrolizat konoplje pripravljen alkalazom ima snažan citotoksičan utjecaj na stanice, dok frakcija hidrolizata konoplje pripravljenog neutrazom manja od 1 kDa usporava rast stanične kulture i značajno odgađa početak log faze rasta. Praćenjem parametara uzgoja (koncentracije stanica, vijabilnosti, koncentracije metabolita) i analizom stanične smrti i staničnog ciklusa, zaključeno je da konopljini hidrolizati djeluju jače citotoksično na proizvodnu staničnu liniju od lanenih hidrolizata, pri čemu dolazi do aktivacije mehanizma stanične smrti.

Abstract (english)

Plant hydrolysates have been investigated as a potential substitute for animal serum, an undesirable supplement of the medium for cultivation of production cell lines. The aim of this study was to investigate the in vitro effect of flax and hemp cake hydrolysates on the growth and viability of CHO DP-12 cell line in suspension culture. Except for cultivation parameters (cell concentration, viability, metabolite concentrations), the mechanism of cytotoxic action of hydrolysates on CHO DP- 12 cells had been assessed. No significant effect on the CHO DP-12 cell line was observed, except for two hemp hydrolysates at a concentration of 2 g Lˉ¹. Unfractionated hemp hydrolyzate prepared by Alkalase has a strong cytotoxic effect, whereas fraction of hemp hydrolyzate prepared by Neutrase smaller than 1 kDa slows down cell growth and significantly delays the beginning of the log phase. Therefore, it was concluded that hemp hydrolysates have a stronger cytotoxic effect on the production cell line CHO DP-12 than flax hydrolysates, whereby the mechanisms of cell death are activated.

• Keywords: hidrolizati lana i konoplje; proizvodna stanična linija; stanični ciklus; stanična smrt; vijabilnost
• Keywords (english): flax and hemp hydrolysates; production cell line; cell cycle; cell death; viability
• Language: croatian
• URN:NBN: urn:nbn:hr:159:002672
• Study programme: Title: Bioprocess Engineering; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra inženjer/inženjerka bioprocesnog inženjerstva (magistar/magistra inženjer/inženjerka bioprocesnog inženjerstva)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2021-10-05 10:08:25