Removal of Serum Lipids and Lipid‐Derived Metabolites to Investigate Breast Cancer Cell Biology

Abstract

The use of cultured cells has been instrumental in studying biochemical, molecular, and cellular processes. The composition of serum that cells are maintained in can have a profound impact on important cellular checkpoints. We have analyzed cell growth and apoptosis in an estrogen receptor positive breast cancer cell line in the presence of serum that have been treated to remove steroids or lipids, as well-described in the literature. We showed that maintaining cells in the presence of charcoal-dextran-treated serum causes reduced growth rate, which can be reversed by the addition of estradiol. Silica-treated-serum also slowed down cell growth and induced apoptosis. In order to investigate the role of lipids in these phenotypes, we investigated the levels of a wide range of lipids in different sera. We showed that silica-treatment significantly depletes phosphatidylcholines and cholesterol. We also show that lipogenesis is stimulated when cells are cultured with silica-treated-serum and this was reversed by the addition of exogenous lipids, which also restored growth rate and apoptosis. Our results show that cultured cells are sensitive to different serum, most likely due to the differences in levels of structural and signaling metabolites present in their growth environment.

PDK4 dictates metabolic resistance to ferroptosis by suppressing pyruvate oxidation and fatty acid synthesis

“…Highglucose and no-glucose DMEM didn't contain glutamate and fatty acid. Animal serum used in cell culture provides lipids to cells (Brovkovych et al, 2019). In some experiments, we added 1, 5, or 25 mM glucose to these commercially available glucose-free DMEM (Thermo Fisher Scientific, 11966025).…”

Lipase mimetic cyclodextrins

“…4D ). 29 This result suggests the usefulness of A6 as a selective lipase mimetic in more complex physiologically relevant settings.…”

Ceramide synthase 2 deletion decreases the infectivity of HIV-1

“…CerS2 is the predominant CerS expressed in the kidney ( 21 , 30 ), and proteomic analyses of HEK293 cells have revealed that CerS2 is expressed at 10-fold higher levels than CerS5, whereas CerS1, CerS3, CerS4, and CerS6 were either not detected or detected at trace levels ( 31 ). Although some ceramides are present in the fetal bovine serum ( 32 ), and Cer can be generated by hydrolysis of SM or glucosylceramide ( Fig. 1 ), the CerS-mediated pathways are major routes of SL production ( 18 , 19 , 20 , 21 ).…”