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Comparison of feeder cells formed from primary or SV40 immortalised mouse embryonic fibroblasts for embryonic stem cell culture
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences, Toxicology.
2015 (English)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

The embryonic stem cell test (EST) is a method first validated by ECVAM in 2003 that uses embryonic stem cells (ESCs) to study embryotoxicity of different compounds classifying them as embryotoxic, weakly embryotoxic and non-embryotoxic by looking at some endpoints as inhibition of formation of beating cardiomyocytes, cytotoxicity in ESCs and cytotoxicity in mouse 3T3 fibroblasts after ten days culture. Usually ESCs are cultured on a feeder layer formed by inactivated primary mouse embryonic fibroblasts (MEFs), which are cells derived from the embryo. To reduce the number of animals sacrificed the present study has been investigating an improvement to the EST that consist in trying to grow ESCs above a feeder layer formed by inactivated primary MEFs infected by SV40 virus, being transformed into immortalized feeders. To compare both types of feeder cells some endpoints such as morphological comparison between them, the time they take to form a confluent feeder layer, expression of stemness markers Nanog, Pou5f, and Sox2 by ESCs cultured on both feeder layers or formation of embryoid bodies formed from these ESCs were analysed. Results were promising in the first stages, as no morphological differences were observed but finally data seem to point that SV40 feeders cannot form a feeder layer as efficient as the one formed by regular feeders. However, this cannot be affirmed, as contamination occurred repeatedly during study and results obtained were not as good as it would have been desirable.

Place, publisher, year, edition, pages
2015.
National Category
Pharmacology and Toxicology
Identifiers
URN: urn:nbn:se:uu:diva-246188OAI: oai:DiVA.org:uu-246188DiVA, id: diva2:792270
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Available from: 2017-01-19 Created: 2015-03-03 Last updated: 2018-01-11Bibliographically approved

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