By Andrei I. Ivanov
This quantity presents a complete selection of classical and leading edge protocols and strategies to check the traditional improvement and physiological features of the gastrointestinal process and to version the commonest digestive diseases. The chapters specialise in different examine issues together with ex vivo structures to check gastrointestinal improvement and capabilities, in vivo imaging of the gastrointestinal tract, isolation and characterization of intestinal immune cells, and animal types of gastrointestinal irritation and melanoma. The Gastrointestinal body structure and illnesses: tools and Protocols book targets large viewers of physiologists, mobilephone and developmental biologists, immunologists, and physician-scientists operating within the box of gastroenterology and beyond. Written within the hugely successful Methods in Molecular Biology series structure, chapters comprise introductions to their respective subject matters, lists of the required fabrics and reagents, step by step, quite simply reproducible laboratory protocols, and pointers on troubleshooting and warding off identified pitfalls.
Highly sensible and obviously written, Gastrointestinal body structure and illnesses: tools and Protocols will serve either professional researchers in addition to rookies to the sphere and should supply a special source and specialist tips to trendy laboratory thoughts built for interpreting common capabilities and ailments of the gastrointestinal tract.
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Extra resources for Gastrointestinal Physiology and Diseases: Methods and Protocols (Methods in Molecular Biology)
Biochem J 443:603–618 3. Lee GY, Kenny PA, Lee EH, Bissell MJ (2007) Three-dimensional culture models of normal and malignant breast epithelial cells. Nat Methods 4:359–365 4. Onuma K, Ochiai M, Orihashi K et al (2013) Genetic reconstitution of tumorigenesis in primary intestinal cells. Proc Natl Acad Sci U S A 110:11127–11132 5. Wang F, Scoville D, He XC et al (2013) Isolation and characterization of intestinal stem cells based on surface marker combinations and colony-formation assay. Gastroenterology 145:383–395 Chapter 3 Co-culture of Gastric Organoids and Immortalized Stomach Mesenchymal Cells Nina Bertaux-Skeirik, Jomaris Centeno, Rui Feng, Michael A.
16 Yoshiaki Maru et al. 2. Dissociate crypts into single cells by adding 1 ml of Accumax to the pellet (see Note 7). Vortex for 3 s and incubate for 5 min at 37 °C in a water bath. 3. Add 5 ml of cold PBS to wash out the enzyme. Centrifuge for 5 min at 400 × g to precipitate single cells and discard the supernatant. 4. Resuspend single cells with 1× organoid culture media. Carefully disperse the cells by pipetting for 5–10 times with P1000 pipetman (see Note 8). 5. Add 80 μl of ice-cold liquid Matrigel into each well of a 12-well plate and incubate at 37 °C for 20 min to completely solidify Matrigel (see Note 9).
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