Project description:Organoid models have been developed for a number of tissues including the liver. Currenthepatic organoid models are generally simplistic, composed of hepatocytes orcholangiocytes, rendering them less physiologically relevant when compared to nativetissue. To address this we have developed an approach that does not require 2Dpatterning, which is ECM independent and growth factor free, to mimic embryonic liverdevelopment that produces massive quantities of liver like organoids. Using single-cell RNAsequencing and immunofluorescence we demonstrate a liver-like cellular repertoire,presenting with vascular luminal structures,and a population of resident macrophage – theKupffer cells. The organoids exhibit key liver functions including drug metabolism, serumprotein production, urea synthesis and coagulation factor production, which preservedpost-translational modifications such as N-glycosylation and functionality. The organoidscan be transplanted and maintained inmice producing human albumin long term.Theorganoids exhibit a complex cellular repertoire reflective of the organ, havede novovascularization, and liver like function. This isa pre-requisite for a myriad of applicationsfrom cellular therapy, tissue engineering, drug toxicity assessment, disease modeling, tobasic developmental biology.
Project description:The lack of physiological parity between 2D cell culture and in vivo culture has led to the development of more organotypic models, such as organoids. Organoid models have been developed for a number of tissues, including the liver. Current organoid protocols are characterized by a reliance on extracellular matrices (ECMs), patterning in 2D culture, costly growth factors and a lack of cellular diversity, structure, and organization. Current hepatic organoid models are generally simplistic and composed of hepatocytes or cholangiocytes, rendering them less physiologically relevant compared to native tissue. We have developed an approach that does not require 2D patterning, is ECM independent, and employs small molecules to mimic embryonic liver development that produces large quantities of liver-like organoids. Using single-cell RNA sequencing and immunofluorescence, we demonstrate a liver-like cellular repertoire, a higher order cellular complexity, presenting with vascular luminal structures, and a population of resident macrophages: Kupffer cells. The organoids exhibit key liver functions, including drug metabolism, serum protein production, urea synthesis and coagulation factor production, with preserved post-translational modifications such as N-glycosylation and functionality. The organoids can be transplanted and maintained long term in mice producing human albumin. The organoids exhibit a complex cellular repertoire reflective of the organ and have de novo vascularization and liver-like function. These characteristics are a prerequisite for many applications from cellular therapy, tissue engineering, drug toxicity assessment, and disease modeling to basic developmental biology.
Project description:Single cell RNA-seq characterization of human induced pluripotent stem cells-derived human cortical organoids (hCO) cultured in control and XG-supplemented conditions Reference: Narazaki, Genta; Miura, Yuki; Pavlov, Sergey D.; Thete, Mayuri Vijay; Roth, Julien G.; Avar, Merve; Kim, Ji-il; Hudacova, Zuzana; Heilshorn, Sarah C.; Pașca, Sergiu P. Scalable production of human cortical organoids using a biocompatible polymer. GSE232581. DOI: (2025).
Project description:The application of organoids has been limited by the lack of methods for producing uniformly mature organoids at scale. This study introduces an organoid culture platform, called UniMat, which addresses the challenges of uniformity and maturity simultaneously. UniMat is designed to not only ensure consistent organoid growth but also facilitate an unrestricted supply of soluble factors by a 3D geometrically-engineered, permeable membrane-based platform. Using UniMat, we demonstrate the scalable generation of kidney organoids with enhanced uniformity in both structure and function compared to conventional methods. Notably, kidney organoids within UniMat matured significantly better, showing increased expression of nephron transcripts, more in vivo-like cell-type balance, and better vascularization. Moreover, UniMat's design offers a more standardized organoid model for drug testing, as demonstrated by its consistent response to a polycystic-kidney-disease drug. In essence, UniMat presents a transformative platform for organoid technology, promising applications in organ development, disease modeling, and drug screening.
Project description:We investigated changes in transcriptome in Egfr KO PSCs co-cultured with pancreatic tumor organoids compared to Egfr WT PSCs co-cultured with the same pancreatic tumor organoids. We also investigated the changes in transcriptome in pancreatic tumor organoids co-cultured with Egfr WT or Egfr KO PSCs.
Project description:We investigated differences in epithelial cells flow-sorted from PDAC tumors derived from the orthotopic transplantation of PDAC T-LOH KPC organoids with either Egfr WT PSCs or Egfr KO PSCs
Project description:RNA-sequencing of flow-sorted Smad4 wild-type (WT) or Smad4 knockout (KO) KvPC (i.e. KRASG12V; p53 mutant) pancreatic ductal adenocarcinoma (PDAC) organoids from monocultures or co-cultures with pancreatic stellate cells (PSCs) and RNA-sequencing of flow-sorted PSCs from the same co-cultures. We investigated changes in transcriptome in Smad4 KO KvPC PDAC organoids compared to Smad4 WT KvPC PDAC organoids in monoculture or in co-culture with PSCs. We also investigated the changes in transcriptome in PSCs co-cultured with Smad4 WT or Smad4 KO KvPC PDAC organoids.