eoe sub articles

EoE Sub Articles

1. Passage of Organoid Cultures
<ol>
	<li>Add 1 ml of cold basal medium to the basement matrix in the well. Using a 1 ml micropipette, vigorously pipette up and down until the basement matrix is broken up. Transfer to a 15 ml tube. Place on ice.</li>
	<li>Take up organoids in the pipette and vigorously pipette 10x up and down. The breaking of the organoids can be observed by the eye.</li>
</ol>
2. Microinjection of Organoid Culture.
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establishing an infection model through microinjection of gastric organoids with pathogenic bacteria

Source: Bartfeld, S. et al. <a href="https://app.jove.com/v/53359">Organoids as Model for Infectious Diseases: Culture of Human and Murine Stomach Organoids and Microinjection of Helicobacter Pylori.</a> J. Vis. Exp.(2015).
This video demonstrates a method to establish a gastric organoid infection model by microinjecting gastric pathogen Helicobacter pylori into the lumen of the organoids. This model helps in studying infection-induced changes in gastric cells.

Establishing an Infection Model Through Microinjection of Gastric Organoids with Pathogenic Bacteria

1. Passage of Organoid Cultures

	Add 1 ml of cold basal medium to the basement matrix in the well. Using a 1 ml micropipette, vigorously pipette up and ...

Under a stereomicroscope, observe gastric organoids, three-dimensional cellular structures composed of differentiated gastric cells, and stem cells.
Introduce a microinjector loaded with Helicobacter pylori, a gastric pathogen.
Insert the needle into the target organoid and inject the bacterial suspension.&#160;
Observe a cloudy solution develop within the lumen.
H.pylori employs a specific adhesin protein on its surface to bind to the gastric cell receptor, establishing a stable interaction between the bacteria and host cells.
This interaction triggers the elongation of the bacterial secretion pilus, a large tunnel-shaped structure that extends toward the host cell membrane.
H.pylori uses the secretion pilus to inject virulence factors, including cytotoxic associated gene A or CagA into the host cell.
Once inside the cell, CagA undergoes phosphorylation by the host cell kinase.
Phosphorylated CagA deregulates multiple intracellular signaling cascades that alter host cell functions, promoting cancerous transformation.&#160;
Collect the infected organoids for further analysis.