PXD010533

PXD010533 is an original dataset announced via ProteomeXchange.

Title	Post-thaw restoration of differentiation and function in cryopreserved human hepatocytesPost-thaw restoration of differentiation and function in cryopreserved human hepatocytes
Description	Isolated human hepatocytes are used in all facets of liver research, from the clinical management of liver failure to in vitro studies of drug disposition. Cryopreservation provides a reliable source of hepatocytes, but the associated cellular stress causes a highly variable and heterogeneous loss of a differentiated phenotype, manifested by a decreased ability to form cell-matrix and cell-cell interactions. We reasoned that this problem could be mitigated at the post-thawing stage, which would increase the availability of well-functioning human hepatocytes. We applied quantitative global proteomics to analyze the differences between attached and non-attached fractions of cryopreserved human hepatocyte batches. Hepatocytes that were unable to attach to a collagen matrix showed many signs of cellular stress, including a glycolytic phenotype and activation of the heat shock response, with increased apoptosis activation as the ultimate consequence. Further analysis of the activated stress pathways revealed an increase in early apoptosis immediately after thawing, hinting at the possibility of stress reversal. Therefore, we transiently treated the cells with compounds aimed at decreasing cellular stress via different mechanisms. We found that brief exposure to the pan-caspase apoptosis inhibitor Z-VAD-FMK restored the ability to attach to collagen, and promoted a differentiated morphology with increased metabolic function. Further, Z-VAD-FMK treatment did not alter hepatocyte protein expression, suggesting that these ‘rescued’ cells would be suitable for applications where hepatocytes of high quality are required. Conclusion: Cryopreserved human hepatocytes are affected by considerable amounts of cellular stress, often resulting in loss of differentiation with a decreased ability to form cell-matrix and cell-cell interactions. This can be alleviated by brief apoptosis inhibition post-thawing, substantially improving key morphological and functional properties of these cells.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_04:47:19.256.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Jacek Wisniewski
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive; LTQ Orbitrap

Revision	Datetime	Status	ChangeLog Entry
0	2018-07-23 05:10:20	ID requested
1	2019-01-02 01:57:41	announced
⏵ 2	2024-10-22 04:47:20	announced	2024-10-22: Updated project metadata.

10.1007/s00204-018-2375-9;

Ö, lander M, Wi, ś, niewski JR, Fl, ö, rkemeier I, Handin N, Urdzik J, Artursson P, A simple approach for restoration of differentiation and function in cryopreserved human hepatocytes. Arch Toxicol, 93(3):819-829(2019) [pubmed]

curator keyword: Biomedical

submitter keyword: Human hepatocyte, attachement, cryopreservation

Jacek R. Wisniewski
contact affiliation	Max-Planck Institute of Biochemistry, Martinsried
contact email	jwisniew@biochem.mpg.de
lab head
Jacek Wisniewski
contact affiliation	Proteomics
contact email	jwisniew@biochem.mpg.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/01/PXD010533

PRIDE project URI

• PRIDE
  1. PXD010533
     1. Label: PRIDE project
     2. Name: Post-thaw restoration of differentiation and function in cryopreserved human hepatocytesPost-thaw restoration of differentiation and function in cryopreserved human hepatocytes