Summary of Study ST004231
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 PR002669. The data can be accessed directly via it's Project DOI: 10.21228/M8NN9R This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST004231 |
| Study Title | Metabolomic studies of H9c2 cells samples |
| Study Summary | Cardiomyocyte differentiation is a complex process involving significant metabolic remodeling, but its impact on cellular redox state and cell damage remains poorly understood. Using metabolomic, biophysical, and biochemical approaches, we characterized, in vitro, the metabolic shift of differentiating cardiomyocytes and its implications for oxidative damage. We found that differentiating cardiomyocytes undergo a broad metabolic reprogramming from a glycolytic to an oxidative state, marked by increased activity in key pathways, including malate-aspartate shuttle, glutathione metabolism, and tricarboxylic acid cycle. This metabolic transition was associated with mitochondrial enlargement and increased reactive oxygen species (ROS) production. Intriguingly, despite ROS increase, differentiated cells maintained similar levels of DNA damage as cardiomyoblasts and were more resistant to a H₂O₂ challenge. Our findings suggest that metabolic adaptations during cardiomyocyte differentiation enhance their capacity to mitigate oxidative stress damage, providing an adaptive avenue that enables cardiomyocyte survival upon exposure to a rich oxygen environment. |
| Institute | University of Campinas |
| Last Name | Amaral |
| First Name | Alan |
| Address | Rua Josué de Castro, s/n - Cidade Universitária, Campinas - SP, 13083-970 |
| a228197@dac.unicamp.br | |
| Phone | +5527996413870 |
| Submit Date | 2025-09-16 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | cdf |
| Analysis Type Detail | GC-MS |
| Release Date | 2025-09-29 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002669 |
| Project DOI: | doi: 10.21228/M8NN9R |
| Project Title: | Quantitative analysis of metabolomics alterations in H9c2 cardiomyocytes during differentiation |
| Project Summary: | Cardiomyocyte differentiation is a complex process involving significant metabolic remodeling, but its impact on cellular redox state and cell damage remains poorly understood. Using metabolomic, biophysical, and biochemical approaches, we characterized, in vitro, the metabolic shift of differentiating cardiomyocytes and its implications for oxidative damage. We found that differentiating cardiomyocytes undergo a broad metabolic reprogramming from a glycolytic to an oxidative state, marked by increased activity in key pathways, including malate-aspartate shuttle, glutathione metabolism, and tricarboxylic acid cycle. This metabolic transition was associated with mitochondrial enlargement and increased reactive oxygen species (ROS) production. Intriguingly, despite ROS increase, differentiated cells maintained similar levels of DNA damage as cardiomyoblasts and were more resistant to a H₂O₂ challenge. Our findings suggest that metabolic adaptations during cardiomyocyte differentiation enhance their capacity to mitigate oxidative stress damage, providing an adaptive avenue that enables cardiomyocyte survival upon exposure to a rich oxygen environment. |
| Institute: | University of Campinas |
| Last Name: | Amaral |
| First Name: | Alan |
| Address: | Rua Josué de Castro, s/n - Cidade Universitária, Campinas - SP, 13083-970 |
| Email: | a228197@dac.unicamp.br |
| Phone: | +5527996413870 |
Subject:
| Subject ID: | SU004383 |
| Subject Type: | Cultured cells |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
Factors:
Subject type: Cultured cells; Subject species: Rattus norvegicus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Time |
|---|---|---|---|
| SA486178 | ENDO_01 | H9c2 cells | T0 |
| SA486179 | ENDO_02 | H9c2 cells | T0 |
| SA486180 | ENDO_03 | H9c2 cells | T0 |
| SA486181 | ENDO_04 | H9c2 cells | T03 |
| SA486182 | ENDO_05 | H9c2 cells | T03 |
| SA486183 | ENDO_06 | H9c2 cells | T03 |
| SA486184 | ENDO_07 | H9c2 cells | T05 |
| SA486185 | ENDO_08 | H9c2 cells | T05 |
| SA486186 | ENDO_09 | H9c2 cells | T05 |
| SA486187 | ENDO_10 | H9c2 cells | T07 |
| SA486188 | ENDO_11 | H9c2 cells | T07 |
| SA486189 | ENDO_12 | H9c2 cells | T07 |
| SA486190 | ENDO_13 | H9c2 cells | T10 |
| SA486191 | ENDO_14 | H9c2 cells | T10 |
| SA486192 | ENDO_15 | H9c2 cells | T10 |
| Showing results 1 to 15 of 15 |
Collection:
| Collection ID: | CO004376 |
| Collection Summary: | H9c2 cells are an established rat-derived cell line from embryonic heart tissue. These immortalized myoblasts can be differentiated into a more cardiac-like phenotype using agents like retinoic acid. Given this and the lack of knowledge about changes in the metabolism of these cells during their differentiation process, commercial immortalized cells were differentiated in medium containing retinoic acid for different periods of time (3, 5, 7, and 10 days) and their internal content was extracted in ice-cold methanol. The samples were stored at -80°C until the derivatization and analysis stage by GC. |
| Sample Type: | H9c2 cells |
Treatment:
| Treatment ID: | TR004392 |
| Treatment Summary: | No treatment. |
Sample Preparation:
| Sampleprep ID: | SP004389 |
| Sampleprep Summary: | Preparation for Drying: 1. From each sample, remove 187 μL and place in a 10 mL falcon tube. Mix the content well using vortex. This mixture is the sample pool for preparing QCs. 2. Aliquot 100 μL of the pool into 14 eppendorf tubes of 1.5 mL. 3. Aliquot 100 μL of the sample into a new 1.5 mL eppendorf tube. 4. Centrifuge the samples at 15,700 g for 20 minutes at 4°C. 5. Remove the supernatant and place in new 1.5 mL eppendorf tubes. Drying and Derivatization 1. Place the eppendorf tubes to dry under vacuum in the SpeedVac using the alcoholic program for 1 hour. (This is an estimated time). 2. Methoxymation: a. Once dried, add 50 μL of MeOX solution (10 mg/mL) to each sample tube. Cap the tube immediately after adding the reagent. Note: perform this step in the fume hood. b. Mix for 30 seconds on vortex. c. Once all tubes are ready, store in darkness for 16 hours. 3. Silylation: a. Add 50 μL of BSTFA (with 1% TMS) to each sample tube. Cap the tube immediately after the reagent is added. b. Mix for 30 seconds on vortex. c. Place the tubes in the heating block at 70°C for 1 hour. d. After this time, remove the tubes and let them cool for 30 minutes. e. Transfer the content of the eppendorf tube to the 200 μL insert (with spring) and complete the volume with 100 μL of methyl stearate standard solution (3 ppm). Gently agitate on vortex. |
Chromatography:
| Chromatography ID: | CH005349 |
| Instrument Name: | Agilent 5975C |
| Column Name: | Agilent HP5-MS (30m x 0.25mm, 0.25 um) |
| Column Temperature: | 60 ºC for 1 min (10 ºC/min) until reach 325 ºC |
| Flow Gradient: | - |
| Flow Rate: | 1.5 mL/min |
| Injection Temperature: | 300 ºC |
| Solvent A: | - |
| Solvent B: | - |
| Oven Temperature: | 60 ºC |
| Transferline Temperature: | 290 ºC |
| Chromatography Type: | GC |
Analysis:
| Analysis ID: | AN007046 |
| Analysis Type: | MS |
| Detector Type: | Triple-Axis Detector (TAD) |
| Chromatography ID: | CH005349 |
| Num Factors: | 5 |
| Num Metabolites: | 51 |
| Units: | Normalized Intensity (a.u.) |
