Summary of Study ST001148
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 PR000767. The data can be accessed directly via it's Project DOI: 10.21228/M8J68Z This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST001148 |
| Study Title | Effect of cell harvesting technique and storage on metabolic profiles in human skin fibroblasts |
| Study Type | Method |
| Study Summary | Human skin fibroblasts were cultured in MEM media supplemented with 15% FBS. Cells were harvested using (i) trypsinization, (ii) scraping, (iii) methanol fixation and scraping, (iV) methanol fixation, scraping, and drying. Targeted metabolomics analysis of organic acids, amino acids, and acycarnitines was conducted at Mayo Clinic Metabolomics Core Facility. |
| Institute | Mayo Clinic |
| Department | Neurology |
| Last Name | Wilkins |
| First Name | Jordan |
| Address | 200 First St SW |
| wilkins.jordan@mayo.edu | |
| Phone | 5072933857 |
| Submit Date | 2019-02-27 |
| Analysis Type Detail | GC-MS/LC-MS |
| Release Date | 2019-09-23 |
| Release Version | 1 |
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Collection:
| Collection ID: | CO001207 |
| Collection Summary: | Prior to harvest, media was removed and cells were rinsed with PBS. For trypsinization, cells were dissociated by incubation with 0.05% trypsin at 37ºC for 5 minutes. Dissociated cells were resuspended in MEM (containing 15% FBS) to neutralize trypsin activity. Cells were centrifuged at 1000 x g for 5 min at 4ºC. Pelleted cells were resuspended in PBS and centrifuged at 1000 x g for 5 min at 4°C. The supernatant was aspirated, cell pellets were flash frozen in liquid nitrogen, and frozen pellets were stored at -80ºC. For scraping, cells were detached in 1 mL PBS using a rubber scraper. Detached cells were centrifuged at 1000 x g for 5 min at 4°C, and PBS was aspirated. Cell pellets were flash frozen in liquid nitrogen and stored at -80ºC. For methanol fixation, 1 mL 80% methanol (pre-chilled on dry ice) was added to cells. Tissue culture plates were rocked by hand for about 20 seconds to evenly distribute methanol solution. Cells were detached in 80% methanol using a rubber scraper. Suspensions were transferred to an Eppendorf tube and stored at -80ºC. For methanol fixation and drying, cells were harvested in 1 mL 80% methanol, and the supernatant was evaporated using a Speed Vacuum at room temperature. Dried samples were stored at -80ºC. Frozen samples were stored at -80ºC for 48 hours, two weeks, or four weeks. |
| Sample Type: | Cultured fibroblasts |
| Collection Frequency: | 48 hours, 2 weeks, and 4 weeks |
| Storage Conditions: | -80℃ |
| Storage Vials: | 1.5 mL Eppendorf tubes |
