Summary of Study ST003704
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 PR002299. The data can be accessed directly via it's Project DOI: 10.21228/M8FZ5G 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.
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| Study ID | ST003704 |
| Study Title | Hyperglycemia leads to BMSC(bone marrow mesenchymal stromal cells) impaired osteogenesis, enhanced adipogenesis, and altered metabolism |
| Study Type | metabolomics |
| Study Summary | Diabetes, a major risk factor for osteoporosis, negatively impacts bone health, but the mechanisms underlying the effects of hyperglycemia on bone marrow mesenchymal stromal cells (BMSC) are not fully understood. This study investigates how high glucose conditions influence BMSC differentiation, proliferation, viability, and metabolism. Our results demonstrate that high glucose inhibits osteogenesis, as evidenced by significantly reduced alkaline phosphatase activity, impaired calcium deposition, and downregulation of key osteogenic genes (RUNX2, Osteocalcin, Osteopontin). Additionally, high glucose conditions promote adipogenesis, characterized by increased lipid accumulation, larger fat droplets, and upregulation of adipogenic genes (PPARγ2, CEBPα, Leptin, AdipoQ), suggesting a shift towards fat cell differentiation. Furthermore, BMSCs cultured in high glucose show decreased proliferation, elevated DNA damage, increased oxidative stress, enhanced apoptosis and senescence, particularly in later passages, highlighting the negative impact of hyperglycemia on BMSC viability. Metabolomic profiling of osteogenic and adipogenic differentiation in normal and high glucose conditions revealed key metabolic shifts, with Nicotinamide Adenine Dinucleotide (NAD+) and L-glutamate/α-ketoglutarate (α-KG) identified as critical metabolites driving these processes. Supplementation with NAD+ and α-KG in high glucose conditions significantly enhanced ALP activity and upregulated osteogenic markers, suggesting that these metabolites play a vital role in restoring osteogenic potential under hyperglycemic conditions. Overall, our findings suggest that high glucose promotes adipogenesis at the expense of osteogenesis, exacerbates cellular damage, and accelerates aging in BMSC. The identification of NAD+ and α-KG as key regulators in this process provides new insights into the metabolic mechanisms behind impaired bone health in diabetes and highlights potential therapeutic avenues to counteract these detrimental effects. Further research into the role of NAD+ and α-KG in BMSC differentiation under high glucose may offer novel strategies for managing diabetes-related bone diseases. |
| Institute | University of Adelaide |
| Department | Health and Medical Sciences |
| Laboratory | MSC Lab |
| Last Name | Shirazi |
| First Name | Suzanna |
| Address | Adelaide University, North Terrace, Adelaide, SA, 5000, Australia |
| suzanna.shirazi@adelaide.edu.au | |
| Phone | 0420245950 |
| Submit Date | 2025-01-20 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-02-24 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002299 |
| Project DOI: | doi: 10.21228/M8FZ5G |
| Project Title: | Metabolomic profiling of human bone marrow mesenchymal stromal stem cells in high glucose |
| Project Type: | Metabolomics |
| Project Summary: | A study on the effect of high glucose level on the differentiation and metabolomic changes of human bone marrow mesenchymal stem cells (BMSC). Human BMSC were cultured under normal and high glucose level media with or without the osteogenic or adipogenic conditions. A range of tests were performed on the cells to look at differentiation, stress response and metabolomics changes in the cells. |
| Institute: | University of Adelaide |
| Department: | Health and Medical Sciences |
| Laboratory: | MSC Lab |
| Last Name: | Shirazi |
| First Name: | Suzanna |
| Address: | Adelaide University, North Terrace, Adelaide, SA, 5000, Australia |
| Email: | suzanna.shirazi@adelaide.edu.au |
| Phone: | 0420245950 |
| Funding Source: | NHMRC |
Subject:
| Subject ID: | SU003836 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample Class | Timepoint | Sample source |
|---|---|---|---|---|
| SA405394 | NDO231_Adipo-Low 21D | Adipogenic Media | 21D | Human BMSC |
| SA405395 | NDO231_Adipo-Hi 21D | Adipogenic Media | 21D | Human BMSC |
| SA405396 | NDO356_Adipo-Low 21D | Adipogenic Media | 21D | Human BMSC |
| SA405397 | NDO356_Adipo-Hi 21D | Adipogenic Media | 21D | Human BMSC |
| SA405398 | NDO362_Adipo-Low 21D | Adipogenic Media | 21D | Human BMSC |
| SA405399 | NDO362_Adipo-Hi 21D | Adipogenic Media | 21D | Human BMSC |
| SA405400 | NDO370_Adipo-Low 21D | Adipogenic Media | 21D | Human BMSC |
| SA405401 | NDO370_Adipo-Hi 21D | Adipogenic Media | 21D | Human BMSC |
| SA405402 | NDO370_Adipo-Hi 7D | Adipogenic Media | 7D | Human BMSC |
| SA405403 | NDO231_Adipo-Low 7D | Adipogenic Media | 7D | Human BMSC |
| SA405404 | NDO356_Adipo-Hi 7D | Adipogenic Media | 7D | Human BMSC |
| SA405405 | NDO362_Adipo-Low 7D | Adipogenic Media | 7D | Human BMSC |
| SA405406 | NDO362_Adipo-Hi 7D | Adipogenic Media | 7D | Human BMSC |
| SA405407 | NDO362_Adipo-Hi 7D DUP | Adipogenic Media | 7D | Human BMSC |
| SA405408 | NDO370_Adipo-Low 7D | Adipogenic Media | 7D | Human BMSC |
| SA405409 | NDO231_Adipo-Hi 7D | Adipogenic Media | 7D | Human BMSC |
| SA405410 | NDO356_Adipo-Low 7D | Adipogenic Media | 7D | Human BMSC |
| SA405411 | NDO370_Cont-Hi 21D | Control Media | 21D | Human BMSC |
| SA405412 | NDO231_Cont-Low 21D | Control Media | 21D | Human BMSC |
| SA405413 | NDO356_Cont-Low 21D | Control Media | 21D | Human BMSC |
| SA405414 | NDO231_Cont-Hi 21D | Control Media | 21D | Human BMSC |
| SA405415 | NDO370_Cont-Low 21D | Control Media | 21D | Human BMSC |
| SA405416 | NDO362_Cont-Hi 21D | Control Media | 21D | Human BMSC |
| SA405417 | NDO362_Cont-Low 21D | Control Media | 21D | Human BMSC |
| SA405418 | NDO356_Cont-Hi 21D | Control Media | 21D | Human BMSC |
| SA405419 | NDO231_Cont-Hi 7D | Control Media | 7D | Human BMSC |
| SA405420 | NDO356_Cont-Low 7D | Control Media | 7D | Human BMSC |
| SA405421 | NDO356_Cont-Hi 7D | Control Media | 7D | Human BMSC |
| SA405422 | NDO362_Cont-Low 7D | Control Media | 7D | Human BMSC |
| SA405423 | NDO362_Cont-Hi 7D | Control Media | 7D | Human BMSC |
| SA405424 | NDO370_Cont-Low 7D | Control Media | 7D | Human BMSC |
| SA405425 | NDO370_Cont-Hi 7D | Control Media | 7D | Human BMSC |
| SA405426 | NDO231_Cont-Low 7D | Control Media | 7D | Human BMSC |
| SA405427 | NDO231_Osteo-Low 21D | Osteogenic Media | 21D | Human BMSC |
| SA405428 | NDO370_Osteo-Hi 21D | Osteogenic Media | 21D | Human BMSC |
| SA405429 | NDO231_Osteo-Hi 21D | Osteogenic Media | 21D | Human BMSC |
| SA405430 | NDO370_Osteo-Low 21D | Osteogenic Media | 21D | Human BMSC |
| SA405431 | NDO362_Osteo-Hi 21D | Osteogenic Media | 21D | Human BMSC |
| SA405432 | NDO362_Osteo-Low 21D | Osteogenic Media | 21D | Human BMSC |
| SA405433 | NDO356_Osteo-Hi 21D | Osteogenic Media | 21D | Human BMSC |
| SA405434 | NDO356_Osteo-Low 21D | Osteogenic Media | 21D | Human BMSC |
| SA405435 | NDO370_Osteo-Hi 7D | Osteogenic Media | 7D | Human BMSC |
| SA405436 | NDO370_Osteo-Low 7D | Osteogenic Media | 7D | Human BMSC |
| SA405437 | NDO362_Osteo-Hi 7D | Osteogenic Media | 7D | Human BMSC |
| SA405438 | NDO362_Osteo-Low 7D | Osteogenic Media | 7D | Human BMSC |
| SA405439 | NDO356_Osteo-Hi 7D | Osteogenic Media | 7D | Human BMSC |
| SA405440 | NDO356_Osteo-Low 7D | Osteogenic Media | 7D | Human BMSC |
| SA405441 | NDO231_Osteo-Low 7D | Osteogenic Media | 7D | Human BMSC |
| SA405442 | NDO231_Osteo-Hi 7D | Osteogenic Media | 7D | Human BMSC |
| Showing results 1 to 49 of 49 |
Collection:
| Collection ID: | CO003829 |
| Collection Summary: | BMSC were isolated from the posterior iliac crests of adult volunteers with informed consent and ethical approval from the Royal Adelaide Hospital. BM mononuclear cells were cultured in α-MEM with 10% FCS, 2 mM L-glutamine, and 100 μM L-ascorbate-2-phosphate. BMSCs were cultured in media with normal (1 g/L) or high (4.5 g/L) glucose and incubated at 37°C with 5% CO2 |
| Sample Type: | Cultured cells |
Treatment:
| Treatment ID: | TR003845 |
| Treatment Summary: | BMSC from 3 human donors were cultured for 7 and 21 days in normal or high glucose DMEM media under control (DMEM cat num # 11885-084 500ml and #11995-065 Thermofisher), osteogenic ( Fetal calf serum 5%, Additive’s 1X, Ascorbate-2P 100µM, Dexamethasone (Na/PO4) 0.1µM, KH2PO4 2.6mM), or adipogenic ( Fetal calf serum 10%, Additive’s 1X, Ascorbate-2P 100µM, Dexamethasone (Na/PO4) 0.1µM, Indomethacin 60µM) conditions. Media was changed every 3 days. A total of 48 samples were processed by Australian Wine Research Institute, South Australia, where 110 compounds were detected using HILIC LC-MS. Compound identification was based on internal and external spectral databases |
Sample Preparation:
| Sampleprep ID: | SP003842 |
| Sampleprep Summary: | 1. 900 uL of a mixture of Methanol and Chloroform (75% methanol, 25% chloroform) were added to the Eppendorf tube containing the cell media (sample). 2. The Eppendorf containing the sample and the solvents was vortexed for 1 minutes and then extracted for 15 minutes using an orbital shaker. 3. After extraction, the Eppendorf tubes were centrifuged at 14,500 rpm for 15 minutes. The supernatant was collected and transferred to a new 2mL Eppendorf tube. 4. The remaining media was submitted to a second extraction using 300 uL of the Methanol Chloroform mixture (described in point 1) 5. Step 2 and 3 were repeated and the supernatant obtained from the second extraction was combined to the one obtained from the first extraction. 6. 500 uL of ice cold MilliQ water was added to the Eppendorf containing the extract. The Eppendorf tube was vortexed for 1 minute and centrifuged at 14,500 rpm for 15 minutes. 7. The supernatant (top layer) was collected, transferred to a glass test tube and dried down using a nitrogen evaporator (using a nitrogen flow of 3.5 ml/min at 25 degrees.) 8. The dried samples were resuspended using 75 uL of Mobile phase A (20 mM Ammonim Carbonate Buffer in MilliQ water) and 25 uL of Mobile Phase B (Acetonitrile) 9. After resuspension the samples (100uL) were transferred in HPLC vial inserts for LC-MS analysis |
Combined analysis:
| Analysis ID | AN006077 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Thermo Vanquish Horizon |
| Column | Waters Xbridge Amide (150 x 2.1mm, 3.5um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Orbitrap ID-X Tribrid |
| Ion Mode | NEGATIVE |
| Units | chromatographic areas with no units |
Chromatography:
| Chromatography ID: | CH004615 |
| Chromatography Summary: | Solvent A: 20 mM Ammonim Carbonate Buffer, pH=9; chrom. flow gradient linearly decreased from 75% B to 50% B from 0.5 to 36min |
| Instrument Name: | Thermo Vanquish Horizon |
| Column Name: | Waters Xbridge Amide (150 x 2.1mm, 3.5um) |
| Column Temperature: | 30°C |
| Flow Gradient: | 0-0.5min: 75% B, 0.5 to 36min: linearly decreased from 75% B to 50% B |
| Flow Rate: | 0.4 mL/min |
| Solvent A: | 100% water; 20 mM Ammonim Carbonate |
| Solvent B: | 100% Acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS005784 |
| Analysis ID: | AN006077 |
| Instrument Name: | Thermo Orbitrap ID-X Tribrid |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Thermo Compound Discoverer Software (version 3.3) was used to perform molecular feature extraction and feature grouping into compounds. The term molecular feature describes a two-dimensional bounded signal: a chromatographic peak (retention time) and a mass spectral peak (m/z). MS1 signals were acquired for the sample replicates and the pooled mix replicates (“PBQC”). The PBQC is a pooled mix of the samples that is used to monitor the instrument performance along the sequence. In addition, 3 replicates of the pooled mix were analysed in MS/MS mode, using HCD fragmentation. These MS/MS analyses were used to provide more reliable putative identifications and structural information of the detected compounds. The raw data matrix was median normalised to correct for any possible inter-run instrument variability. Following this, the CV% among the Pooled Mix replicates was calculated and compounds showing a CV% value above 25 were removed from the data matrix because inconsistent through the analysis batch. The first 3 PBQC replicates were not included in the CV calculation because they were used to condition the instrument. The total number of detected compounds was 109. |
| Ion Mode: | NEGATIVE |
