dc.contributor.author | Messner, Catherine | |
dc.contributor.author | Babrak, Lmar | |
dc.contributor.author | Titolo, Gaia | |
dc.contributor.author | Caj, Michaela | |
dc.contributor.author | Miho, Enkelejda | |
dc.contributor.author | Suter-Dick, Laura | |
dc.date.accessioned | 2022-03-28T12:19:49Z | |
dc.date.available | 2022-03-28T12:19:49Z | |
dc.date.issued | 2021-04-22 | |
dc.identifier.doi | 10.3390/ijms22094372 | |
dc.identifier.uri | https://irf.fhnw.ch/handle/11654/33410 | |
dc.identifier.uri | http://dx.doi.org/10.26041/fhnw-4152 | |
dc.description.abstract | 3D 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.iso | en_US | en_US |
dc.publisher | MDPI | en_US |
dc.relation.ispartof | International Journal of Molecular Sciences | en_US |
dc.rights | Attribution 3.0 United States | * |
dc.rights.uri | http://creativecommons.org/licenses/by/3.0/us/ | en_US |
dc.accessRights | Anonymous | * |
dc.subject | Single cell sequencing | en_US |
dc.subject | In vitro | en_US |
dc.subject | Liver | en_US |
dc.subject | Liver fibrosis | en_US |
dc.subject | Liver microtissues | en_US |
dc.title | Single Cell Gene Expression analysis in a 3D microtissue liver model reveals cell type-specific responses to pro-fibrotic TGF-β1 stimulation | en_US |
dc.type | 01 - Zeitschriftenartikel, Journalartikel oder Magazin | * |
dc.volume | 22 | en_US |
dc.issue | 9 | en_US |
dc.spatial | Basel | en_US |
fhnw.publicationState | Published | en_US |
fhnw.ReviewType | Anonymous ex ante peer review of a complete publication | en_US |
fhnw.InventedHere | Yes | en_US |
fhnw.IsStudentsWork | no | en_US |
fhnw.openAccessCategory | Gold | |