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Differentiation of Mesenchymal Stem Cells and Cell Line 3T3-L1K into Adipocytes as an In vitro Toxicological Model
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Pharmaceutical Biosciences.
2016 (English)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

In vitro toxicological models are required to assess toxicity outside of animals. In this report focus has been put on introducing pre-adipocyte cell line 3T3-L1K as a model to asses effects caused by phospholipidsosis inducing compounds and potential pro-adipogenic endocrine disturbing chemicals by differentiating the cells into adipocytes and looking at potential inhibitory/inducing effects.

Phospholipidosis inducing compound have been reported to inhibit differentiation of monocytes and dendritic cells in vitro, possibly by disrupting mechanisms and signalling required for cells to differentiate. Cells were treated with these compounds and assessed by looking at gene expression for marker genes of adipogenesis, methylation patterns on a gene regulator of adipogenesis (peroxisome proliferator activated receptor γ 2) and from a functional imaging assay to look at lipid droplet formation and morphological differences in the samples.

Furthermore a few selected endocrine disturbing substances on their effect on adipogenesis were evaluated on the same assays.

The results presented show statistical evidence for 2 phospholipid accumulating compounds to cause inhibition of 5 marker genes involved in adipogenesis on murine cell line 3T3-L1K in a concentration dependent fashion. Results from another differentiation with other known phospholipidosis inducing compounds showed similar inhibitory trends. Imaging of cells gave similar results, where it was visible that cells treated with phospholipid accumulating compounds had inhibition on both amount of lipid droplets and adipocyte phenotype.

Presented optimised protocol for endocrine disturbing substances could detect change in adipogenic markers for bisphenol A, which were undetectable in standard in vitro chemically induced differentiation normal differentiation.

Place, publisher, year, edition, pages
Keyword [en]
phospholipidosis, adipogenesis, endocrine disturbing chemicals, 3T3-L1K
National Category
Other Natural Sciences
URN: urn:nbn:se:uu:diva-274317OAI: oai:DiVA.org:uu-274317DiVA: diva2:896228
External cooperation
Subject / course
Educational program
Master of Science Programme in Pharmacy
Available from: 2016-10-06 Created: 2016-01-20 Last updated: 2016-10-06Bibliographically approved

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