Summary of Study ST000167
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000145. The data can be accessed directly via it's Project DOI: 10.21228/M87596 This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
Study ID | ST000167 |
Study Title | Cold Storage of Rat Hepatocyte Spheroids |
Study Type | Storage condition testing |
Study Summary | The purpose of this study was to optimize conditions for cold storage of rat hepatocyte spheroids without freezing. Rat hepatocytes were isolated by a two-step perfusion method; hepatocyte spheroids were formed during 48 h of rocked culture in serum-free medium (SFM). Spheroids were then maintained in rocked culture at 37°C (control condition) or cold stored at 4°C for 24 or 48 h in six different cold storage solutions: SFM alone; SFM + 1 mM deferoxamine (Def); SFM + 1 ?M cyclosporin A (CsA); SFM + 1 mM Def + 1 ?M CsA, University of Wisconsin (UW) solution alone, UW + 1 mM Def. |
Institute | Mayo Clinic |
Department | Division of Experimental Surgery |
Last Name | Nyberg |
First Name | Scott |
Nyberg.scott@mayo.edu | |
Submit Date | 2015-05-14 |
Num Groups | 9 |
Total Subjects | 45 |
Raw Data Available | No |
Analysis Type Detail | LC-MS |
Release Date | 2015-06-28 |
Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Collection:
Collection ID: | CO000172 |
Collection Summary: | Hepatocytes were isolated from male SpragueDawley rats (300400 g; Harlan, Indianapolis, IN, USA) by a two-step perfusion method as previously described (33). All harvests yielded hepatocytes with viability exceeding 95% by trypan blue dye exclusion. Freshly isolated hepatocytes were suspended in SFM, composed of Williams E supplemented with 0.2 U/ml insulin, 6 ?g/ml transferrin, 100 U/ml penicillin G, 100 mg/ml streptomycin, 3 g/L human albumin, 2.2 g/L sodium bicarbonate, 1 g/L L-carnitine, 2.0 mM L-glutamine, 100 nM dexamethasone, 40 ng/ml glucagon, 20 ng/ml Gly-His-Lys, 1,000 U/L heparin, 1 mg/L warfarin, and 5 ng/ml of mouse epidermal growth factor (EGF) (3). The cells, suspended in SFM, were placed in a spheroid box (33 × 28 × 6 cm) custom-made of polycarbonate by Mayo Division of Engineering and siliconized with Sigmacote for 30 min (20) and gently rocked continuously at a frequency of 10 cycles per minute (0.17 Hz) to induce spheroid formation and to maintain spheroids in suspension. Final hepatocyte concentration was 5 × 105 cells/ml per spheroid box. All culture conditions were maintained in a 5% CO2, 37°C incubator as previously described (20). Spheroids were centrifuged and resuspended in fresh culture media every 24 h. |
Sample Type: | Liver |