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 |
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Sample Preparation:
Sampleprep ID: | SP000186 |
Sampleprep Summary: | After 48 h of continuous rocking, the spheroids were washed with SFM at room temperature. The culture medium was changed to the cold storage medium (UW alone, UW + 1 mM Def; SFM alone; SFM + 1 mM Def; SFM + 1 mM Def + 1 ?M CsA; SFM + 1 ?M CsA) at room temperature. Spheroids were left in rocked culture without cold storage as control. Spheroids intended for cold storage were rocked in glass dishes (10 × 8 × 2 cm) custom-made by Mayo Division of Engineering and siliconized with Sigmacote for 30 min and then transferred into 50-ml conical tubes (BD Falcon; 1 × 106cells/ml, 20 ml in total) and put on ice for 1 h. Of note, spheroid box and glass dishes differ in size (i.e., volume capacity) but possess similar properties of spheroid formation and culture. Finally, tubes of spheroids were placed in a refrigerator to be cold stored at 4°C. After 24 or 48 h of cold storage treatment, the spheroids were centrifuged at 50 × g for 5 min, and the supernatant fluid was removed. Warm SFM (20 ml) at 37°C was added to each tube. The tubes were mixed gently prior to adding 20 ml of cell suspension to each glass culture dish and continued to rocking culture. Cultures were maintained in a humidified incubator at 37°C with a 5% CO2 atmosphere. |