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Adapting Cancer Cell Lines to a Fully Defined Medium

Rafnsdóttir, Ólöf Birna (2019) MOBN03 20182
Degree Projects in Molecular Biology
Popular Abstract
Lately, there has been a world-wide increase in the awareness of animal cruelty e.g. in the entertainment business, meat industry, in the scientific community, and among people in general. This has called for a scrutiny of how we treat animals and in the scientific community it has stimulated research to reduce animal experiment. Experimental systems usually considered animal cruelty free, like cell culturing, should be on the frontier of totally animal free experiments, however it is not and unfortunately many researchers using cell culture do not ponder over this. In order to grow cells outside the body, special cell culture media are needed to provide nutrients for the cells. Cell medium traditionally contains serum, the most common... (More)
Lately, there has been a world-wide increase in the awareness of animal cruelty e.g. in the entertainment business, meat industry, in the scientific community, and among people in general. This has called for a scrutiny of how we treat animals and in the scientific community it has stimulated research to reduce animal experiment. Experimental systems usually considered animal cruelty free, like cell culturing, should be on the frontier of totally animal free experiments, however it is not and unfortunately many researchers using cell culture do not ponder over this. In order to grow cells outside the body, special cell culture media are needed to provide nutrients for the cells. Cell medium traditionally contains serum, the most common being foetal bovine serum (FBS), a cocktail of different known and unknown compounds, obtained from blood harvested via cardiac puncture of the foetus of female cows on their way to slaughter. This possibly inflicts pain and suffering to the unborn animal. This is not the only problem scientists face using serum as a growth supplement. There are variations in compound compositions and concentrations between batches of serum. Implying that different batches of serum could give different results in experiment conducted in the exact same manner. The primary aim of this study was to compose a totally defined medium (DM) based on the literature and experience in the laboratory and then adapt cancer cell lines to this medium, and compare with cells grown in medium containing serum. This study focused on adapting the human breast cancer cell line JIMT-1 and the human pancreatic cancer cell line MiaPaCa-2 to grow in DM. The adaption process, gradually increasing the DM in relation to the serum containing medium took 2 months. Then different experiments were preformed to evaluate how well the cells thrived in the DM. Growth curves were used to determine the population doubling time. Flow cytometry methods were used to determine protein expression. Dose response experiments were performed to investigate the effects of the experimental cancer stem cell inhibiting compound salinomycin. The JIMT-1 cells and the MiaPaCa-2 cells thrived in the DM and the parameters investigated were similar to those obtained when the cells were cultured in medium supplemented with serum. However, morphologically, the cells were more rounded in the DM compared to the medium supplemented with serum and they were not attached as firm as in the later medium. Growing cells in 2D is not very physiological as cells grow in 3D in the body. Thus, JIMT-1 and MiaPaCa-2 cells were seeded in an animal product free 3D culture system. Both cell types thrived in the total animal free 3D cell culture system.

Although both the JIMT-1 and MiaPaCa-2 cells seem to have been adapted to DM further research has to be done to confirm the present work and also to investigate why there are changes in morphology and what impact that has. (Less)
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author
Rafnsdóttir, Ólöf Birna
supervisor
organization
course
MOBN03 20182
year
type
H2 - Master's Degree (Two Years)
subject
language
English
id
8995075
date added to LUP
2019-09-16 13:53:28
date last changed
2019-09-16 13:53:28
@misc{8995075,
  author       = {{Rafnsdóttir, Ólöf Birna}},
  language     = {{eng}},
  note         = {{Student Paper}},
  title        = {{Adapting Cancer Cell Lines to a Fully Defined Medium}},
  year         = {{2019}},
}