Molecular and Functional Characterization of Human Intestinal Organoids and Monolayers for Modeling Epithelial Barrier

Patient-derived organoid models can transform drug discovery for inflammatory bowel disease, but differentiation and functional characterization inconsistencies limit that transformation. Jelinsky et al. profiled molecular and cellular features across a range of intestinal organoid models using bulk RNA sequencing, among other technologies, and examined differentiation and establishing a functional epithelial barrier.

BMP gradient along the intestinal villus axis controls zonated enterocyte and goblet cell states

Beumer et al. show that the bone morphogenetic protein (BMP) signaling pathway controls the creation of functional zones along the crypt-villus axis. A gene lipid uptake gene signature is enriched in the villus tip and could be therapeutically exploited with BMP inhibitors.

CD127+ CD94+ innate lymphoid cells expressing granulysin and perforin are expanded in patients with Crohn’s disease

The researchers designate two distinct innate lymphoid cell subtypes. Then, using single-cell RNA sequencing and further characterization methods, one subtype was found to express toxic compounds granulysin and perforin in the intestines of Chron’s disease patients.

Collection of cells for single-cell RNA sequencing using high-resolution fluorescence microscopy

Segeren et al. describe a workflow to capture, image, and collect fluorescent human retina pigment epithelium cells for SORT-seq using the VYCAP puncher system. This protocol is relevant for cells that cannot be FACS-sorted.

Identification of Enteroendocrine Regulators by Real-Time Single-Cell Differentiation Mapping

This paper presents Neurog3Chrono, a bi-fluorescent reporter for the differentiation process of enteroendocrine cells, the intestines’ hormone-excreting cells. It measures fluorescence in time and per cell, enabling the real-time arrangement of single-cell gene expression profiles.