Vpliv tripsinizacije na celice Caco-2

Cuculić, Nika (author), Oven, Irena (mentor)

Caco-2 celična linija se uporablja v številnih predkliničnih raziskavah. Ena izmed najbolj izraženih lastnosti celične linije Caco-2 je njihova zmožnost spontane diferenciacije. Zaradi različnega rokovanja s celično linijo Caco-2 med laboratoriji, v kar spada tudi uporaba različnih koncentracij tripsina ter čas izpostavljenosti celic tripsinu, se rezultati raziskav med laboratoriji lahko zelo razlikujejo in niso primerljivi. Preverili smo vpliv različnih koncentracij tripsina na celično viabilnost, prilagajanje celic na površino ter izražanje določenih genov z metodo verižne reakcije s polimerazo v realnem času z namenom, da bi ocenili vpliv tripsina na Caco-2 celično linijo ter v kakšni meri tripsin vpliva na razlike med rezultati, pridobljenimi med različnimi laboratoriji, in s pomočjo rezultatov izdelali univerzalen protokol, ki bi pomagal tudi drugim raziskovalcem, ki uporabljajo celično linijo Caco-2.

• Language: Slovenian
• Keywords: tripsinizacija, celična viabilnost, prilagajanje celic, izražanje genov, qPCR
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [N. Cuculić]
• Year: 2018
• PID: 20.500.12556/RUL-99492
• UDC: 576.33:577.2(043.2)
• COBISS.SI-ID: 8908409
• Publication date in RUL: 28.01.2018

Secondary language

• Language: English
• Title: Impact of trypsinization on Caco-2 cells

Abstract

Caco-2 cell line is widely used in different preclinical and clinical studies. One of the most prominent features of Caco-2 cell line is their ability of spontaneous differentiation. Due to different handling procedures of Caco-2 cell line, which include the use of different concentrations and variable time of the treatmeant with trypsin, results differ between laboratories which makes comparsion impossible. We tested the impact of different concentrations of trypsin on cell viability, attachment of the cells on surface and the expression of different genes with quantitave polymerase chain reaction method with purpose of evaluating the impact of trypsin on Caco-2 cell line and to what extent it affects the differences in results between laboratories. With the help of results we could make a standardized protocol which could help other scientists dealing with Caco-2 cell line.

• Keywords: trypsinization, cell viability, cell adaption, gene expression, PCR