Colonic Crypt Isolation
Colonic Crypt Isolation - Related Videos
04:37
Intestinal Crypts Isolation: A Method to Isolate Whole Crypts from Small Intestine of Murine Model
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2023
In this video, we demonstrate a technique to isolate whole crypts from a mouse’s small intestine. The isolated whole crypts can be used to propagate long-lived and self-renewing 3D organoids in vitro.
06:57
Generation of Murine Primary Colon Epithelial Monolayers from Intestinal Crypts
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Cited by 12 •
2021
In this protocol, we describe how to generate murine primary epithelial colon monolayers directly from intestinal crypts. We provide experimental approaches to generate confluent monolayers on permeable filters, confluent monolayers for scratch wound healing and biochemical studies, and sparse and confluent monolayers for immunofluorescence analysis.
07:53
Quantification of Colonic Stem Cell Mutations
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Cited by 1 •
2015
We report significant improvements for the reproducible measurement of somatic colonic stem cell mutations after exposure of mice to potential DNA damaging agents.
06:59
Isolation of Primary Myofibroblasts from Mouse and Human Colon Tissue
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Cited by 43 •
2013
The myofibroblast is an influential stromal cell of the gastrointestinal tract that regulates important physiologic processes in both normal and disease states. We describe a technique that allows for the isolation of primary myofibroblasts from both mouse and human colon tissue, which can be utilized for in vitro experimentation.
06:16
Cryosectioning Method for Microdissection of Murine Colonic Mucosa
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Cited by 5 •
2015
Here we describe a simple method for the isolation of spatially distinct murine colonic epithelial surface cells from the underlying crypt-base cells.
FAQs
Calcium chelation weakens calcium-dependent adhesion between cells and their surrounding tissue, making the epithelial units easier to release. This chemical step reduces the need for forceful disruption, which helps preserve crypt structure during subsequent processing. Maintaining that organization is important when researchers want to examine morphology, gene expression, stem cell activity, or treatment responses in material derived from the colon.
Gentle mechanical agitation helps release crypts after adhesion has been weakened, while filtration separates the liberated units from larger tissue fragments. The balance between these steps matters because excessive disruption can damage the structures being studied, whereas insufficient agitation may leave crypts embedded in tissue. Filtration then enriches the preparation for downstream examination.
Intact crypts retain epithelial organization, allowing investigators to study structural features together with molecular and functional responses. This format supports examination of morphology, gene expression, stem cell activity, and reactions to experimental treatments within a preserved tissue unit. Such information can be more informative for epithelial changes than observations made after organization has been lost.
Comparing healthy and tumor-associated crypt preparations helps identify differences linked to colorectal cancer biology. The approach can reveal early epithelial changes and distinguish tissue-associated patterns that may not be apparent from a general tissue sample. These comparisons provide a way to investigate how organization, gene expression, stem cell activity, or treatment responses differ across disease-related contexts.
The workflow begins with tissue processing and calcium chelation to disrupt adhesion, followed by gentle mechanical agitation to release crypts. The resulting material is passed through filtration to remove larger fragments and enrich the isolated units. Researchers can then examine the preparation directly for morphology, gene expression, stem cell activity, or responses to experimental treatments.
Researchers can evaluate several outcome types from the isolated preparation, including crypt morphology, gene expression, stem cell activity, and responses to experimental treatments. These measurements connect structural observations with epithelial biology and treatment effects. In cancer research, the resulting data can support analysis of early changes and comparisons between non-tumor-associated and tumor-associated tissue.
Isolated crypts support studies of colorectal cancer biology by providing material for comparisons between healthy and tumor-associated tissue. They also contribute to ex vivo models, which allow researchers to investigate epithelial behavior and responses to experimental treatments outside the original tissue context. This makes the technique relevant to studying disease-associated changes and evaluating treatment-related effects.