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dc.contributor.authorMessner, Catherine
dc.contributor.authorBabrak, Lmar
dc.contributor.authorTitolo, Gaia
dc.contributor.authorCaj, Michaela
dc.contributor.authorMiho, Enkelejda
dc.contributor.authorSuter-Dick, Laura
dc.date.accessioned2022-03-28T12:19:49Z
dc.date.available2022-03-28T12:19:49Z
dc.date.issued2021-04-22
dc.identifier.doi10.3390/ijms22094372
dc.identifier.urihttps://irf.fhnw.ch/handle/11654/33410
dc.identifier.urihttp://dx.doi.org/10.26041/fhnw-4152
dc.description.abstract3D cell culture systems are widely used to study disease mechanisms and therapeutic interventions. Multicellular liver microtissues (MTs) comprising HepaRG, hTERT-HSC and THP-1 maintain multicellular interactions and physiological properties required to mimic liver fibrosis. However, the inherent complexity of multicellular 3D-systems often hinders the discrimination of cell type specific responses. Here, we aimed at applying single cell sequencing (scRNA-seq) to discern the molecular responses of cells involved in the development of fibrosis elicited by TGF-β1. To obtain single cell suspensions from the MTs, an enzymatic dissociation method was optimized. Isolated cells showed good viability, could be re-plated and cultured in 2D, and expressed specific markers determined by scRNA-seq, qRT-PCR, ELISA and immunostaining. The three cell populations were successfully clustered using supervised and unsupervised methods based on scRNA-seq data. TGF-β1 led to a fibrotic phenotype in the MTs, detected as decreased albumin and increased αSMA expression. Cell-type specific responses to the treatment were identified for each of the three cell types. They included HepaRG damage characterized by a decrease in cellular metabolism, prototypical inflammatory responses in THP-1s and extracellular matrix remodeling in hTERT-HSCs. Furthermore, we identified novel cell-specific putative fibrosis markers in hTERT-HSC (COL15A1), and THP-1 (ALOX5AP and LAPTM5).en_US
dc.language.isoen_USen_US
dc.publisherMDPIen_US
dc.relation.ispartofInternational Journal of Molecular Sciencesen_US
dc.rightsAttribution 3.0 United States*
dc.rights.urihttp://creativecommons.org/licenses/by/3.0/us/en_US
dc.accessRightsAnonymous*
dc.subjectSingle cell sequencingen_US
dc.subjectIn vitroen_US
dc.subjectLiveren_US
dc.subjectLiver fibrosisen_US
dc.subjectLiver microtissuesen_US
dc.titleSingle Cell Gene Expression analysis in a 3D microtissue liver model reveals cell type-specific responses to pro-fibrotic TGF-β1 stimulationen_US
dc.type01 - Zeitschriftenartikel, Journalartikel oder Magazin*
dc.volume22en_US
dc.issue9en_US
dc.spatialBaselen_US
fhnw.publicationStatePublisheden_US
fhnw.ReviewTypeAnonymous ex ante peer review of a complete publicationen_US
fhnw.InventedHereYesen_US
fhnw.IsStudentsWorknoen_US
fhnw.openAccessCategoryGold


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