PXD014971-1

PXD014971 is an original dataset announced via ProteomeXchange.

Title	Are These Cardiomyocytes? Protocol Development Reveals Impact of Sample Preparation on the Accuracy of Identifying Cardiomyocytes by Flow Cytometry
Description	Several protocols now support efficient differentiation of human pluripotent stem cells to cardiomyocytes (hPSC-CMs) but these still indicate line-to-line variability. As the number of studies implementing this technology expands, accurate assessment of cell identity is paramount to well-defined studies that can be replicated among laboratories. While flow cytometry is apt for routine assessment, a standardized protocol for assessing cardiomyocyte identity has not yet been established. Therefore, the current study leveraged targeted mass spectrometry to confirm the presence of troponin proteins in day 25 hPSC-CMs and systematically evaluated multiple anti-troponin antibodies and sample preparation protocols for their suitability in assessing cardiomyocyte identity. Results demonstrate challenges to interpreting data generated by published methods and inform the development of a robust protocol for routine assessment of hPSC-CMs. The data, workflow for antibody evaluation, and standardized protocol described here should benefit investigators new to this field and those with expertise in hPSC-CM differentiation.
HostingRepository	PanoramaPublic
AnnounceDate	2019-08-12
AnnouncementXML	Submission_2019-08-12_09:44:16.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Matthew Waas
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	Label:13C(6)15N(2); Label:13C(6)15N(4)
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2019-08-09 15:55:20	ID requested
⏵ 1	2019-08-12 09:44:18	announced

Waas M, Weerasekera R, Kropp EM, Romero-Tejeda M, Poon EN, Boheler KR, Burridge PW, Gundry RL, Are These Cardiomyocytes? Protocol Development Reveals Impact of Sample Preparation on the Accuracy of Identifying Cardiomyocytes by Flow Cytometry. Stem Cell Reports, 12(2):395-410(2019) [pubmed]

submitter keyword: quality control, flow cytometry, troponin, cardiomyocytes, mass spectrometry, standard operating protocol

Rebekah Gundry
contact affiliation	Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI
contact email	rgundry@mcw.edu
lab head
Matthew Waas
contact affiliation	Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI
contact email	mwaas@mcw.edu
dataset submitter

Panorama Public dataset URI