Summary of Study ST002070
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 PR001312. The data can be accessed directly via it's Project DOI: 10.21228/M8499N 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 | ST002070 |
Study Title | Lipidomic Comparison of 2D and 3D Colon Cancer Cell Culture Models |
Study Type | Untargeted Lipidomics |
Study Summary | Altered lipid metabolism is one of the hallmarks of cancer. Cellular proliferation and de novo synthesis of lipids are related to cancer progression. In this study, we evaluated the lipidomic profile of two-dimensional (2D) monolayer and multicellular tumor spheroids from the HCT 116 colon carcinoma cell line. We utilized serial trypsinization on the spheroid samples to generate three cellular populations representing the proliferative, quiescent, and necrotic regions of the spheroid. This analysis enabled a comprehensive identification and quantification of lipids produced in each of the spheroid layer and 2D cultures. We show that lipid subclasses associated with lipid droplets form in oxygen-restricted and acidic regions of spheroids and are produced at higher levels than in 2D cultures. Additionally, sphingolipid production, which is implicated in cell death and survival pathways, is higher in spheroids relative to 2D cells. Finally, we show that increased numbers of lipids comprised of polyunsaturated fatty acids (PUFAs) are produced in the quiescent and necrotic regions of the spheroid. The lipidomic signature for each region and cell culture type highlights the importance of understanding the spatial aspects of cancer biology. These results provide additional lipid biomarkers in the tumor microenvironment that can be further studied during potential therapeutic studies which target pivotal lipid production pathways. |
Institute | The Ohio State University |
Last Name | Tobias |
First Name | Fernando |
Address | 100 W 18th Avenue, Columbus, Ohio, 43210, USA |
tobias.62@osu.edu | |
Phone | 12345678907 |
Submit Date | 2022-01-30 |
Raw Data Available | Yes |
Raw Data File Type(s) | d |
Analysis Type Detail | LC-MS |
Release Date | 2022-02-14 |
Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
Project ID: | PR001312 |
Project DOI: | doi: 10.21228/M8499N |
Project Title: | Lipidomic Comparison of 2D and 3D Colon Cancer Cell Culture Models |
Project Type: | Untargeted Lipidomics |
Project Summary: | Altered lipid metabolism is one of the hallmarks of cancer. Cellular proliferation and de novo synthesis of lipids are related to cancer progression. In this study, we evaluated the lipidomic profile of two-dimensional (2D) monolayer and multicellular tumor spheroids from the HCT 116 colon carcinoma cell line. We utilized serial trypsinization on the spheroid samples to generate three cellular populations representing the proliferative, quiescent, and necrotic regions of the spheroid. This analysis enabled a comprehensive identification and quantification of lipids produced in each of the spheroid layer and 2D cultures. We show that lipid subclasses associated with lipid droplets form in oxygen-restricted and acidic regions of spheroids and are produced at higher levels than in 2D cultures. Additionally, sphingolipid production, which is implicated in cell death and survival pathways, is higher in spheroids relative to 2D cells. Finally, we show that increased numbers of lipids comprised of polyunsaturated fatty acids (PUFAs) are produced in the quiescent and necrotic regions of the spheroid. The lipidomic signature for each region and cell culture type highlights the importance of understanding the spatial aspects of cancer biology. These results provide additional lipid biomarkers in the tumor microenvironment that can be further studied during potential therapeutic studies which target pivotal lipid production pathways. |
Institute: | The Ohio State University |
Department: | Department of Chemistry and Biochemistry |
Laboratory: | Amanda B Hummon Research Group |
Last Name: | Tobias |
First Name: | Fernando |
Address: | 100 W 18th Avenue, Columbus, Ohio, 43210, USA |
Email: | tobias.62@osu.edu |
Phone: | 1234567890 |
Subject:
Subject ID: | SU002152 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Male |
Cell Strain Details: | HCT 116 |
Subject Comments: | HCT 116 cell line was cultured as a 2D monolayer and 3D spheroids |
Species Group: | Mammals |
Factors:
Subject type: Cultured cells; Subject species: Homo sapiens (Factor headings shown in green)
mb_sample_id | local_sample_id | GROUP | TYPE |
---|---|---|---|
SA194712 | 23 2D1 | 2D1_2 | 2D |
SA194713 | 41 2D1 | 2D1_3 | 2D |
SA194714 | 5 2D1 | 2D1 | 2D |
SA194715 | 9 2D2 | 2D2_1 | 2D |
SA194723 | 27 2D2 | 2D_2_2 | 2D |
SA194716 | 45 2D2 | 2D2_3 | 2D |
SA194717 | 14 2D3 | 2D3_1 | 2D |
SA194718 | 32 2D3 | 2D3_2 | 2D |
SA194719 | 50 2D3 | 2D3_3 | 2D |
SA194720 | 18 2D4 | 2D4_1 | 2D |
SA194721 | 36 2D4 | 2D4_2 | 2D |
SA194722 | 54 2D4 | 2D4_3 | 2D |
SA194724 | 8 P1C | Core1_1 | SPHEROID |
SA194735 | 26 P1C | Core_1_2 | SPHEROID |
SA194725 | 44 P1C | Core1_3 | SPHEROID |
SA194726 | 12 P2C | Core2_1 | SPHEROID |
SA194727 | 30 P2C | Core2_2 | SPHEROID |
SA194728 | 48 P2C | Core2_3 | SPHEROID |
SA194729 | 17 P3C | Core3_1 | SPHEROID |
SA194730 | 35 P3C | Core3_2 | SPHEROID |
SA194731 | 53 P3C | Core3_3 | SPHEROID |
SA194732 | 21 P4C | Core4_1 | SPHEROID |
SA194733 | 39 P4C | Core4_2 | SPHEROID |
SA194734 | 57 P4C | Core4_3 | SPHEROID |
SA194736 | 7 P1M | Middle1_1 | SPHEROID |
SA194737 | 25 P1M | Middle1_2 | SPHEROID |
SA194738 | 43 P1M | Middle1_3 | SPHEROID |
SA194739 | 11 P2M | Middle2_1 | SPHEROID |
SA194740 | 29 P2M | Middle2_2 | SPHEROID |
SA194741 | 47 P2M | Middle2_3 | SPHEROID |
SA194742 | 16 P3M | Middle3_1 | SPHEROID |
SA194743 | 34 P3M | Middle3_2 | SPHEROID |
SA194744 | 52 P3M | Middle3_3 | SPHEROID |
SA194745 | 20 P4M | Middle4_1 | SPHEROID |
SA194746 | 38 P4M | Middle4_2 | SPHEROID |
SA194747 | 56 P4M | Middle4_3 | SPHEROID |
SA194748 | 6 P1O | Outer1_1 | SPHEROID |
SA194749 | 42 P1O | Outer1_3 | SPHEROID |
SA194750 | 10 P2O | Outer2_1 | SPHEROID |
SA194751 | 28 P2O | Outer2_2 | SPHEROID |
SA194752 | 46 P2O | Outer2_3 | SPHEROID |
SA194759 | 24 P1O | Outer_2 | SPHEROID |
SA194753 | 15 P3O | Outer3_1 | SPHEROID |
SA194754 | 33 P3O | Outer3_2 | SPHEROID |
SA194755 | 51 P3O | Outer3_3 | SPHEROID |
SA194756 | 19 P4O | Outer4_1 | SPHEROID |
SA194757 | 37 P4O | Outer4_2 | SPHEROID |
SA194758 | 55 P4O | Outer4_3 | SPHEROID |
SA194760 | 59 QC20 | QC10 | QC |
SA194761 | 60 QC40 | QC11 | QC |
SA194762 | 2 QC10 | QC1 | QC |
SA194763 | 3 QC20 | QC2 | QC |
SA194764 | 4 QC40 | QC3 | QC |
SA194765 | 13 QC10 | QC4 | QC |
SA194766 | 22 QC20 | QC5 | QC |
SA194767 | 31 QC40 | QC6 | QC |
SA194768 | 40 QC20 | QC7 | QC |
SA194769 | 49 QC40 | QC8 | QC |
SA194770 | 58 QC10 | QC9 | QC |
Showing results 1 to 59 of 59 |
Collection:
Collection ID: | CO002145 |
Collection Summary: | Cell Lysates from 2D Monolayers and serially-trypsinized spheroids was subjected to lipid extraction |
Sample Type: | Cultured cells |
Storage Conditions: | -80℃ |
Collection Tube Temp: | 4 |
Additives: | 1X PBS |
Treatment:
Treatment ID: | TR002164 |
Treatment Summary: | HCT 116 2D monolayers were cultured per ATCC protocol, while HCT 116 3D spheroids were cultured for 14 days. At Day 14, the spheroids were subjected to Serial Trypsinization in order to obtain discrete cell populations from spheroids representing the outer, middle, and core regions. |
Sample Preparation:
Sampleprep ID: | SP002158 |
Sampleprep Summary: | Lipid extraction by MTBE/methanol/water solvent system |
Processing Storage Conditions: | On ice |
Extraction Method: | liquid-liquid extraction |
Extract Storage: | -80℃ |
Sample Resuspension: | 9:1 Methanol/Toluene |
Sample Spiking: | equiSPLASH LIPIDOMIX standards |
Combined analysis:
Analysis ID | AN003374 | AN003375 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | Reversed phase | Reversed phase |
Chromatography system | Agilent 1260 | Agilent 1260 |
Column | Thermo Accucore C30 (100 x 2.1mm,2.6um) | Thermo Accucore C30 (100 x 2.1mm,2.6um) |
MS Type | ESI | ESI |
MS instrument type | QTOF | QTOF |
MS instrument name | Agilent 6545 QTOF | Agilent 6545 QTOF |
Ion Mode | POSITIVE | NEGATIVE |
Units | pmol lipid/ ug protein | pmol lipid/ ug protein |
Chromatography:
Chromatography ID: | CH002494 |
Instrument Name: | Agilent 1260 |
Column Name: | Thermo Accucore C30 (100 x 2.1mm,2.6um) |
Column Temperature: | 50 |
Flow Rate: | 400uL/min |
Injection Temperature: | 4 |
Internal Standard: | equiSPLASH LIPIDOMIX |
Sample Injection: | 2 |
Solvent A: | 50% water/50% acetonitrile; 10mM ammonium formate |
Solvent B: | 90% isopropanol/10% acetonitrile; 10 mM ammonium formate |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS003141 |
Analysis ID: | AN003374 |
Instrument Name: | Agilent 6545 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | MS-Only on biological samples, Auto-MSMS on Pooled Sample |
Ion Mode: | POSITIVE |
MS ID: | MS003142 |
Analysis ID: | AN003375 |
Instrument Name: | Agilent 6545 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | MS-Only on biological samples, Auto-MSMS on Pooled Sample |
Ion Mode: | NEGATIVE |