Usporedna istraživanja dinamike rasta CHO-K1 i CCO stanica u kulturi

master's thesis

• Title (english): Comparison of growth parameters of CHO-K1 and CCO cells in culture
• Author: Ivana Porobić
• Mentor: Zlatko Kniewald (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: 2007-12-20, Croatia
• Scientific / art field, discipline and subdiscipline: BIOTECHNICAL SCIENCES; Biotechnology

Abstract

Tehnologija životinjskih stanica te mogućnost njihove primjene u industriji i proizvodnji korisnih proizvoda zauzima značajno mjesto u biotehnologiji i srodnim znanstvenim područjima. Cilj rada je utvrditi parametre uzgoja CCO i CHO-K1 stanične linije uz naglasak na potrošnju i nastajanje metabolita u mediju za uzgoj. Istraživan je rast CCO stanica tijekom 9 dana uzgoja u dva različita medija, DMEM (ATCC) + 10% FBSa i D-MEM/F12 (Gibco) + 10% NCS-a. Utvrđen je veći prirast CCO stanica nakon 5 dana uzgoja u DMEM mediju s 10% FBS-a u usporedbi s uzgojem u mediju s 5% FBS-a i 2,5% FBS-a. Tijekom 5 dana uzgoja CCO i CHO-K1 stanica praćen je utrošak metabolita u mediju. Rezultati pokazuju da CCO stanice ne koriste glukozu kao primarni izvor energije. Mogući izvor energije za ove stanice su aminokiseline, naročito glutamin. Također je utvrđeno nakupljanje manje količine laktata kao metaboličkog nusproizvoda. Određivanje metabolita u D-MEM/F12 mediju pokazuje da CHO-K1 stanice koriste glukozu kao glavni izvor energije, pri čemu se laktat nakuplja u većoj količini. To upućuje na nepotpunu razgradnju glukoze. Ionskom izmjenom u mediju za uzgoj CCO stanica određeno je nakupljanje alanina i glicina, kao i potrošnja treonina, serina, metionina, arginina, taurina i glutamina. Određivanje aminokiselina u mediju za uzgoj CHO-K1 stanica pokazuje nakupljanje alanina i ornitina, kao i potrošnju asparaginske kiseline, glutaminske kiseline, glutamina, serina i treonina. Nakupljanje laktata i alanina u mediju za uzgoj CHO-K1 stanica ukazuje na veliku potrošnju glukoze i aminokiselina što predstavlja nesrazmjer u potrošnji metabolita i energetskim potrebama stanica u kulturi.

Abstract (english)

Application of animal cell technology for production of active biological components has an important place in biotechnology. Object of this study was determination of growth parameters of CCO and CHO-K1 animal cells as well as monitoring of medium metabolite consumption and production. This study reports on growth of CCO cell line during 9 days of cultivation in two different media, DMEM (ATCC) + 10% FBS and D-MEM/F12 (Gibco) + 10% NCS. Growth of CCO cell line was found better in DMEM medium with 10% FBS than in medium with 5% or 2,5% FBS. Consumption of medium metabolites was monitored during 5 days of growth of CCO and CHO-K1 cell lines. The results show that CCO cells do not use glucose as a primary energy source. Possible source of energy for these cells may be amino acids, especially glutamine. However accumulation of lactate as metabolic byproduct was also detected. Metabolite detection of D-MEM/F12 medium showed that CHO-K1 cells use glucose as a main source of energy while lactate accumulates as metabolic byproduct. This indicates of incomplete break-down of glucose. Accumulation of alanine and glycine, as well as consumption of treonine, serine, methionine, arginine, taurine and glutamine in medium for CCO cells cultivation was detected by ion-exchange chromatography. The amino acid detection in medium for CHO-K1 cell cultivation showed accumulation of alanine and ornithine, as well as consumption of asparartate, glutamic acid, glutamine, serine and threonine. Accumulation of lactate and alanine in growth medium for CHO-K1 cells is due to significant glucose and amino acids consumption, which represents typical metabolic disfunction in energy requirement of cultured cells.

• Keywords: aminokiseline; CCO; CHO-K1; metabolizam; stanične kulture
• Keywords (english): amino acids; CCO; cell culture; CHO-K1; metabolism
• Language: croatian
• URN:NBN: urn:nbn:hr:159:563338
• Study programme: Title: Biotechnology; specializations in: Biochemical-Microbiology; Course: Biochemical-Microbiology; Study programme type: university; Study level: graduate; Academic / professional title: diplomirani inženjer biotehnologije; smjer biokemijsko-mikrobiološki (diplomirani inženjer biotehnologije; smjer biokemijsko-mikrobiološki)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Access restricted to students and staff of home institution
• Created on: 2017-09-05 12:04:11