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.

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Study IDST000167
Study TitleCold Storage of Rat Hepatocyte Spheroids
Study TypeStorage condition testing
Study SummaryThe 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
DepartmentDivision of Experimental Surgery
Last NameNyberg
First NameScott
EmailNyberg.scott@mayo.edu
Submit Date2015-05-14
Num Groups9
Total Subjects45
Raw Data AvailableNo
Analysis Type DetailLC-MS
Release Date2015-06-28
Release Version1
Scott Nyberg Scott Nyberg
https://dx.doi.org/10.21228/M87596
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000145
Project DOI:doi: 10.21228/M87596
Project Title:Cold Storage of Rat Hepatocyte Spheroids
Project Summary:Cell-based therapies for liver disease rely on a high-quality supply of hepatocytes and a means for storage during transportation from site of isolation to site of usage. Unfortunately, frozen cryopreservation is associated with unacceptable loss of hepatocyte viability after thawing. 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. Performance metrics after cold storage included viability, gene expression, albumin production, and functional activity of cytochrome P450 enzymes and urea cycle proteins. We observed that cold-induced injury was reduced significantly by the addition of the iron chelator (Def) to both SFM and UW solution. Performance metrics (ammonia detoxification, albumin production) of rat hepatocyte spheroids stored in SFM + Def for 24 h were significantly increased from SFM alone and approached those in control conditions, while performance metrics after cold storage in SFM alone or cold storage for 48 h were both significantly reduced. A serum-free medium supplemented with Def allowed hepatocyte spheroids to tolerate 24 h of cold storage with less than 10% loss in viability and functionality. Further research is warranted to optimize a solution for extended cold storage of hepatocyte spheroids.
Institute:Mayo Clinic
Department:Division of Experimental Surgery
Last Name:Nyberg
First Name:Scott
Email:Nyberg.scott@mayo.edu
Funding Source:This work was funded by grants from NIH (RO1-DK56733), the Marriott Foundation, the Wallace H. Coulter Foundation, the 12th Five-Year National Science and Technology Major Project for Infectious Diseases, China (No. 2012 ZX 10002004-005, Hongling Liu), and Jiangsu province government, China (Yue Yu). The authors declare no conflicts of interest.
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