#METABOLOMICS WORKBENCH jcorstvet3_20250326_115849 DATATRACK_ID:5792 STUDY_ID:ST003849 ANALYSIS_ID:AN006326 PROJECT_ID:PR002407
VERSION             	1
CREATED_ON             	April 8, 2025, 12:50 pm
#PROJECT
PR:PROJECT_TITLE                 	Metabolomic Screening of Biomaterials for Mesenchymal stem/stromal cells (MSC)
PR:PROJECT_TITLE                 	Culture
PR:PROJECT_SUMMARY               	Mesenchymal stem/stromal cells (MSCs) have transformative healthcare potential,
PR:PROJECT_SUMMARY               	but to achieve therapeutic cell numbers, MSCs must be culture expanded. On
PR:PROJECT_SUMMARY               	tissue culture poly(styrene), however, MSC expansion is constrained by
PR:PROJECT_SUMMARY               	replicative senescence, and cells experience progressive loss of therapeutic
PR:PROJECT_SUMMARY               	potency with continued expansion. The lack of large numbers of reliably potent
PR:PROJECT_SUMMARY               	therapeutic cells is considered one of the most critical roadblocks to the
PR:PROJECT_SUMMARY               	success of MSC clinical trials. However, screening of improved MSC culture
PR:PROJECT_SUMMARY               	conditions has been hampered by the fact that there are no characterization
PR:PROJECT_SUMMARY               	tools that might allow prediction of cell fitness early in culture. Metabolism
PR:PROJECT_SUMMARY               	is the most dynamic level of cellular response, and metabolomics may enable the
PR:PROJECT_SUMMARY               	identification of early signatures associated with optimal cell product quality.
PR:PROJECT_SUMMARY               	Therefore, it is hypothesized that early metabolomics data from MSCs cultured on
PR:PROJECT_SUMMARY               	biomaterials will allow prediction of cell fitness later in culture and thus
PR:PROJECT_SUMMARY               	promote development of materials to promote a desired cell phenotype during
PR:PROJECT_SUMMARY               	expansion. The objective of this application is to determine the relationship
PR:PROJECT_SUMMARY               	between biomaterial carrier properties, cell metabolism, and maintenance of MSC
PR:PROJECT_SUMMARY               	fitness during expansion. This objective will be approached through the
PR:PROJECT_SUMMARY               	following specific aims: 1) Evaluate the effects of altering ligand type and
PR:PROJECT_SUMMARY               	stiffness of the biomaterial substrate on metabolism and replicative senescence
PR:PROJECT_SUMMARY               	of human MSCs during expansion in serum, and 2) Evaluate the effects of the
PR:PROJECT_SUMMARY               	substrate heparin content and pattern of initial seeding on metabolism and
PR:PROJECT_SUMMARY               	replicative senescence of human MSCs during expansion in serum-free conditions.
PR:PROJECT_SUMMARY               	The proposed work is innovative because it uses early cellular metabolic
PR:PROJECT_SUMMARY               	responses to design material substrates that will promote cell expansion without
PR:PROJECT_SUMMARY               	senescence in a range of media compositions. Results from these studies are
PR:PROJECT_SUMMARY               	expected to have an important positive impact because they will lead to more
PR:PROJECT_SUMMARY               	efficacious expansion protocols for MSCs and thus produce more effective
PR:PROJECT_SUMMARY               	cell-based therapies for a wide variety of diseases.
PR:INSTITUTE                     	Georgia Institute of Technology
PR:LAST_NAME                     	Corstvet
PR:FIRST_NAME                    	Joseph
PR:ADDRESS                       	North Avenue, Atlanta, GA, 30332, USA
PR:EMAIL                         	jcorstvet3@gatech.edu
PR:PHONE                         	2626410497
#STUDY
ST:STUDY_TITLE                   	Metabolomic Screening of Biomaterials for Mesenchymal stem/stromal cells (MSC)
ST:STUDY_TITLE                   	Culture
ST:STUDY_SUMMARY                 	Mesenchymal stem/stromal cells (MSCs) have transformative healthcare potential,
ST:STUDY_SUMMARY                 	but to achieve therapeutic cell numbers, MSCs must be culture expanded. On
ST:STUDY_SUMMARY                 	tissue culture poly(styrene), however, MSC expansion is constrained by
ST:STUDY_SUMMARY                 	replicative senescence, and cells experience progressive loss of therapeutic
ST:STUDY_SUMMARY                 	potency with continued expansion. The lack of large numbers of reliably potent
ST:STUDY_SUMMARY                 	therapeutic cells is considered one of the most critical roadblocks to the
ST:STUDY_SUMMARY                 	success of MSC clinical trials. However, screening of improved MSC culture
ST:STUDY_SUMMARY                 	conditions has been hampered by the fact that there are no characterization
ST:STUDY_SUMMARY                 	tools that might allow prediction of cell fitness early in culture. Metabolism
ST:STUDY_SUMMARY                 	is the most dynamic level of cellular response, and metabolomics may enable the
ST:STUDY_SUMMARY                 	identification of early signatures associated with optimal cell product quality.
ST:STUDY_SUMMARY                 	Therefore, it is hypothesized that early metabolomics data from MSCs cultured on
ST:STUDY_SUMMARY                 	biomaterials will allow prediction of cell fitness later in culture and thus
ST:STUDY_SUMMARY                 	promote development of materials to promote a desired cell phenotype during
ST:STUDY_SUMMARY                 	expansion. The objective of this application is to determine the relationship
ST:STUDY_SUMMARY                 	between biomaterial carrier properties, cell metabolism, and maintenance of MSC
ST:STUDY_SUMMARY                 	fitness during expansion. This objective will be approached through the
ST:STUDY_SUMMARY                 	following specific aims: 1) Evaluate the effects of altering ligand type and
ST:STUDY_SUMMARY                 	stiffness of the biomaterial substrate on metabolism and replicative senescence
ST:STUDY_SUMMARY                 	of human MSCs during expansion in serum, and 2) Evaluate the effects of the
ST:STUDY_SUMMARY                 	substrate heparin content and pattern of initial seeding on metabolism and
ST:STUDY_SUMMARY                 	replicative senescence of human MSCs during expansion in serum-free conditions.
ST:STUDY_SUMMARY                 	The proposed work is innovative because it uses early cellular metabolic
ST:STUDY_SUMMARY                 	responses to design material substrates that will promote cell expansion without
ST:STUDY_SUMMARY                 	senescence in a range of media compositions. Results from these studies are
ST:STUDY_SUMMARY                 	expected to have an important positive impact because they will lead to more
ST:STUDY_SUMMARY                 	efficacious expansion protocols for MSCs and thus produce more effective
ST:STUDY_SUMMARY                 	cell-based therapies for a wide variety of diseases. From this study, we found
ST:STUDY_SUMMARY                 	that various triglycerides were elevated in older/highly senescent MSC, with
ST:STUDY_SUMMARY                 	longer chain lengths and more unsaturations in the fatty acid chains
ST:STUDY_SUMMARY                 	specifically correlating to higher senescence. We also observed an increased
ST:STUDY_SUMMARY                 	abundance of trihexosylceramides in higher senescent MSC samples.
ST:INSTITUTE                     	Georgia Institute of Technology
ST:LAST_NAME                     	Corstvet
ST:FIRST_NAME                    	Joseph
ST:ADDRESS                       	North Avenue, Atlanta, GA, 30332, USA
ST:EMAIL                         	jcorstvet3@gatech.edu
ST:PHONE                         	2626410497
#SUBJECT
SU:SUBJECT_TYPE                  	Human
SU:SUBJECT_SPECIES               	Homo sapiens
SU:TAXONOMY_ID                   	9606
#SUBJECT_SAMPLE_FACTORS:         	SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day0-Rep1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day0-Rep1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day0-Rep2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day0-Rep2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day0-Rep3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day0-Rep3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day0-Rep4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day0-Rep4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day2-Rep1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 2 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day2-Rep1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day2-Rep2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 2 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day2-Rep2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day2-Rep3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 2 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day2-Rep3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day2-Rep4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 2 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day2-Rep4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day4-Rep1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day4-Rep1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day4-Rep2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day4-Rep2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day4-Rep3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day4-Rep3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day4-Rep4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day4-Rep4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day6-Rep1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 6 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day6-Rep1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day6-Rep2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 6 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day6-Rep2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day6-Rep3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 6 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day6-Rep3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day6-Rep4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 6 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day6-Rep4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day8-Rep1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day8-Rep1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day8-Rep2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day8-Rep2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day8-Rep3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day8-Rep3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day8-Rep4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day8-Rep4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day10-Rep1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 10 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day10-Rep1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day10-Rep2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 10 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day10-Rep2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day10-Rep3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 10 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day10-Rep3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day10-Rep4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 10 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day10-Rep4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day12-Rep1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day12-Rep1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day12-Rep2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day12-Rep2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day12-Rep3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day12-Rep3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	RB182-Day12-Rep4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB182-Day12-Rep4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day0-Rep1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day0-Rep1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day0-Rep2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day0-Rep2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day0-Rep3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day0-Rep3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day0-Rep4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day0-Rep4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day2-Rep1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 2 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day2-Rep1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day2-Rep2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 2 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day2-Rep2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day2-Rep3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 2 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day2-Rep3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day2-Rep4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 2 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day2-Rep4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day4-Rep1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day4-Rep1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day4-Rep2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day4-Rep2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day4-Rep3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day4-Rep3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day4-Rep4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day4-Rep4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day6-Rep1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 6 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day6-Rep1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day6-Rep2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 6 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day6-Rep2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day6-Rep3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 6 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day6-Rep3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day6-Rep4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 6 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day6-Rep4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day8-Rep1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day8-Rep1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day8-Rep2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day8-Rep2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day8-Rep3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day8-Rep3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day8-Rep4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day8-Rep4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day10-Rep1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 10 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day10-Rep1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day10-Rep2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 10 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day10-Rep2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day10-Rep3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 10 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day10-Rep3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day10-Rep4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 10 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day10-Rep4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day12-Rep1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day12-Rep1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day12-Rep2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day12-Rep2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day12-Rep3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day12-Rep3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	RB310-Day12-Rep4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 1 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=RB310-Day12-Rep4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-RGD-pA-1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB182-RGD-pA-1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-RGD-pA-2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB182-RGD-pA-2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-RGD-pA-3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB182-RGD-pA-3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-RGD-pA-4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB182-RGD-pA-4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-RGD-pB-1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB182-RGD-pB-1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-RGD-pB-2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB182-RGD-pB-2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-RGD-pB-3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB182-RGD-pB-3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-RGD-pC-1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 12 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB182-RGD-pC-1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-RGD-pC-2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 12 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB182-RGD-pC-2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-RGD-pC-3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 12 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB182-RGD-pC-3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-Day-0-1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-Day-0-1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-Day-0-2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-Day-0-2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-Day-0-3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-Day-0-3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-Day-0-4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-Day-0-4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-pA-1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-pA-1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-pA-2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-pA-2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-pA-3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-pA-3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-pA-4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-pA-4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-pB-1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-pB-1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-pB-2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-pB-2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-pB-3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-pB-3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-pB-4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-pB-4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-pC-1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-pC-1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-pC-2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-pC-2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 1	C2-RB182-TCP-pC-3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB182-TCP-pC-3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-RGD-pA-1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB310-RGD-pA-1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-RGD-pA-2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB310-RGD-pA-2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-RGD-pA-3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB310-RGD-pA-3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-RGD-pA-4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB310-RGD-pA-4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-RGD-pB-1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB310-RGD-pB-1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-RGD-pB-2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB310-RGD-pB-2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-RGD-pB-3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB310-RGD-pB-3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-RGD-pB-4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB310-RGD-pB-4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-RGD-pC-1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 12 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB310-RGD-pC-1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-RGD-pC-2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 12 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB310-RGD-pC-2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-RGD-pC-3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 12 | Culture substrate:Hydrogel	RAW_FILE_NAME(Raw file name)=C2-RB310-RGD-pC-3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-Day-0-1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-Day-0-1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-Day-0-2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-Day-0-2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-Day-0-3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-Day-0-3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-Day-0-4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 0 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-Day-0-4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-pA-1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-pA-1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-pA-2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-pA-2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-pA-3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-pA-3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-pA-4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 4 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-pA-4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-pB-1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-pB-1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-pB-2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-pB-2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-pB-3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-pB-3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-pB-4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 8 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-pB-4.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-pC-1	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-pC-1.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-pC-2	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-pC-2.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-pC-3	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-pC-3.mzML
SUBJECT_SAMPLE_FACTORS           	Donor 2	C2-RB310-TCP-pC-4	Sample source:Bone Marrow-MSC cells | Cohort:Cohort 2 | Culture day:Day 12 | Culture substrate:TCP	RAW_FILE_NAME(Raw file name)=C2-RB310-TCP-pC-4.mzML
#COLLECTION
CO:COLLECTION_SUMMARY            	Bone marrow MSC collected from healthy adult donors were purchased from
CO:COLLECTION_SUMMARY            	RoosterBio. These cells were expanded for 2 passages before cryopreservation in
CO:COLLECTION_SUMMARY            	CS10 freezing medium (Cryostor) until the study began.
CO:SAMPLE_TYPE                   	Bone marrow
#TREATMENT
TR:TREATMENT_SUMMARY             	Cells were culture expanded and revived from frozen storage in tissue culture
TR:TREATMENT_SUMMARY             	plastic (TCP) flasks with DMEM media, FBS, and antibiotic/antimycotic and seeded
TR:TREATMENT_SUMMARY             	in 6-well culture plates for 3 passages of 4 days each. Cells were harvested
TR:TREATMENT_SUMMARY             	every 2-4 days for 12 days total.
#SAMPLEPREP
SP:SAMPLEPREP_SUMMARY            	Lipids were extracted using cold IPA. Cells were lysed with 50 micron glass
SP:SAMPLEPREP_SUMMARY            	beads and IPA supernatant was removed. This extract was dried and stored at -80C
SP:SAMPLEPREP_SUMMARY            	until day of sample analysis, when it was reconstituted in 3:1 IPA:H20
SP:PROCESSING_STORAGE_CONDITIONS 	-80℃
SP:EXTRACT_STORAGE               	-80℃
#CHROMATOGRAPHY
CH:CHROMATOGRAPHY_TYPE           	Reversed phase
CH:INSTRUMENT_NAME               	Thermo Vanquish
CH:COLUMN_NAME                   	Thermo Accucore C30 (150 x 2.1mm,2.6um)
CH:SOLVENT_A                     	40% water/60% acetonitrile; 10mM ammonium formate; 0.1% formic acid
CH:SOLVENT_B                     	90% isopropanol/10% acetonitrile; 10mM ammonium formate; 0.1% formic acid
CH:FLOW_GRADIENT                 	0.4 mL/min: 0-1.0min: 80%A 20%B, 1.0-5.0min: 40%A 60%B, 5.0-5.5min: 30%A 70%B,
CH:FLOW_GRADIENT                 	5.5-8.0min: 15%A 85%B, 8.0-8.2min: 10%A 90%B, 8.2-10.7min: 100%B, 10.7-12.0min:
CH:FLOW_GRADIENT                 	80%A 20%B.
CH:FLOW_RATE                     	0.4 mL/min
CH:COLUMN_TEMPERATURE            	50
CH:TARGET_SAMPLE_TEMPERATURE     	4
#ANALYSIS
AN:ANALYSIS_TYPE                 	MS
#MS
MS:INSTRUMENT_NAME               	Thermo Orbitrap ID-X Tribrid
MS:INSTRUMENT_TYPE               	Orbitrap
MS:MS_TYPE                       	ESI
MS:ION_MODE                      	POSITIVE
MS:MS_COMMENTS                   	Cohort 2 Data collected at 120,000 FWHM resolution using Thermo Xcalibur
MS:MS_COMMENTS                   	software. QC sample injections were performed after every 8 sample injections to
MS:MS_COMMENTS                   	correct for instrument drift and ensure quality of the run. MS/MS was performed
MS:MS_COMMENTS                   	on top features using iterative DDA on QC sample to assist with feature
MS:MS_COMMENTS                   	annotation. Peak alignment, blank subtraction, QC correction, and feature
MS:MS_COMMENTS                   	annotations were all performed using Compound Discoverer v3.3.
MS:MS_RESULTS_FILE               	ST003849_AN006326_Results.txt	UNITS:peak area	Has m/z:Yes	Has RT:Yes	RT units:Minutes
#END