Language
English
Publication Date
6-28-2024
Journal
JoVE: Journal of Visualized Experiments
DOI
10.3791/66749
PMID
39007612
PMCID
PMC11987515
PubMedCentral® Posted Date
4-11-2025
PubMedCentral® Full Text Version
Author MSS
Abstract
Single cell transcriptomics has revolutionized our understanding of the cell biology of the human body. State-of-the-art human small intestinal organoid cultures provide ex vivo model systems that bridge the gap between animal models and clinical studies. The application of single cell transcriptomics to human intestinal organoid (HIO) models is revealing previously unrecognized cell biology, biochemistry, and physiology of the GI tract. The advanced single cell transcriptomics platforms use microfluidic partitioning and barcoding to generate cDNA libraries. These barcoded cDNAs can be easily sequenced by next generation sequencing platforms and used by various visualization tools to generate maps. Here, we describe methods to culture and differentiate human small intestinal HIOs in different formats and procedures for isolating viable cells from these formats that are suitable for use in single-cell transcriptional profiling platforms. These protocols and procedures facilitate the use of small intestinal HIOs to obtain an increased understanding of the cellular response of human intestinal epithelium at the transcriptional level in the context of a variety of different environments.
Keywords
Humans, Organoids, Intestine, Small, Single-Cell Analysis, Intestinal Mucosa, Gene Expression Profiling, Transcriptome
Published Open-Access
yes
Recommended Citation
Bomidi, Carolyn; Zeng, Xi-Lei; Poplaski, Victoria; et al., "Using Human Intestinal Organoids to Understand the Small Intestine Epithelium at the Single Cell Transcriptional Level" (2024). Faculty and Staff Publications. 4700.
https://digitalcommons.library.tmc.edu/baylor_docs/4700