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MB Sample ID: SA184054
| Local Sample ID: | 3 |
| Subject ID: | SU002030 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
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Collection:
| Collection ID: | CO002023 |
| Collection Summary: | All samples extracted from mice were collected in accordance with regulations and established guidelines for humane treatment of research animals and were reviewed and approved by an Institutional Animal Care and Use Committee, Project Code HD-EX00066. Primary hepatocytes were isolated from male mice between 12-18 weeks of age by the two-step collagenase perfusion method. Mice were fasted 16 hours before the ex-periments. After isolation, cells were plated in M199 media with 10% FBS for 4 hours in 6-well plates pre-coated with collagen I. After cells attached to the plates, they were washed with glucose output media (GOM) (118 mM NaCl, 4.7 mM KCl, 1.2 mM MgSO4, 1.2 mM KH2PO4, 1.2 mM CaCl2, 20 mM NaCO3, 25 mM HEPES pH 7.4, and 0.025% BSA), and incubated in fresh GOM for 2 hour. GOM media was replaced with fresh, pre-warmed GOM media and cellular treatments initiated. Hepatocytes were treated with 5mM unlabeled glutamine for 60 min. Hepatocytes were washed with ice cold PBS twice and immediately frozen in liquid nitrogen. |
| Sample Type: | Liver |
