Volume 17 Issue 12, December 2022

Stem cell models for the trophoblast lineage have been lacking. This protocol describes the reprogramming of human fibroblasts into induced trophoblast stem cells, alongside steps for their molecular and functional characterization.

The authors describe a technique for reprogramming human somatic cells into induced trophoblast stem cells and the conversion of human naive and extended pluripotent stem cells into converted trophoblast stem cells, with functional assays for their differentiation potential.

This two-stage bioreactor-mediated approach for culturing previously uncultured microorganisms involves enrichment of indigenous organisms by using a continuous-flow down-flow hanging sponge bioreactor and subsequent selective batch cultivation.

The authors provide a guide to using the Kraken suite for metagenomics analysis, including classification, quantification and visualization, illustrated by quantification of species in the microbiome and identification of pathogens in a clinical sample.

This protocol provides an enzyme-free method to achieve targeted DNA demethylation based on steric hindrance by dCas9 bound at or close to CpG sites of interest to identify DNA methylation marks causal to altered gene expression.

Inhibitors for carbohydrate digestive enzymes are used to manage conditions like diabetes. In this assay for digestive enzyme activity, sugars are quantified by using high-performance anion-exchange chromatography with pulsed amperometric detection.

The Braille Display valving platform described in this protocol can be used to generate droplets (each with 100 cells), assay reagents and a variety of drug combinations or to sort droplets on the basis of fluorescence signals.

Little is known about the brain mechanisms of hallucinations. This protocol describes the use of a robotic device and sensorimotor method to reproducibly induce and measure presence hallucinations by behavioral assay and fMRI.

Direct carbonization of zeolitic imidazolate framework-type metal-organic frameworks, as described in this protocol, results in nanoporous carbons that can be used for diverse electrochemical applications.

Multispectral large-scale single-cell resolution 3D imaging allows up to eight fluorophores to be captured in a single acquisition. This protocol enables the visualization and exploration of large intact tissue volumes.

The cell membrane turnover kinetics of the main glial glutamate transporter GLT1 are quantified by fusing it to a pH-sensitive fluorescent protein and performing measurements using whole-cell fluorescence recovery after photobleaching.