Which of the following statements is incorrect?

1. Human intestinal organoids are generally passaged as fragments using GCDR.
2. Trypsin-EDTA is required to break up the organoid fragments into single cells before seeding onto transwell inserts.
3. Corning® Matrigel® is used for coating cultureware for the 2D differentiation protocol.
4. The organoid suspension culture protocol is the best way to scale up organoids for monolayers.
5. Once fully differentiated in IntestiCult™ ODM, human intestinal organoids can be passaged and cryopreserved in CryoStor® CS10.

Regarding the key advantages of organoid technologies, which of the following statement is incorrect? (select one):

1. Organoids allow you to study cell type-specific functions, depending on the origin tissue they are derived from.
2. Organoids have comparable performance to traditional cell lines for modeling function, toxicity, and drug responses.
3. Organoids can be maintained long term.
4. Organoids grow as self-organizing “units” with different cell types.

Which of the following statements is correct?

1. Organoid-derived intestinal monolayers (2D) are a useful tool for applications requiring access to the apical surface.
2. Organoid-derived intestinal monolayers (2D) are more physiologically relevant than commonly used in vitro intestinal cell lines.
3. Organoids (3D) and organoid-derived monolayers (2D) can recapitulate the genotype and phenotype of healthy and diseased donors, whereas immortalized intestinal cell lines cannot model genetic diversity within a population.
4. All of the above

Select whether the following statement is true or false:

Cultured intestinal 3D organoids incorporate many of the physiologically relevant features of in vivo intestinal tissue. Although the epithelium is polarized, the lumen is not considered to be fully functional (i.e. no mucus secretion, no ion transport, etc.)

Match the following workflow steps with the correct product:

1. Expansion of human intestinal organoids
2. Differentiation of human intestinal organoids (3D)
3. Generation of human intestinal monolayers (2D)
4. Cryopreservation of human intestinal organoids

1. CryoStor® CS10
2. IntestiCult™ Organoid Growth Media (OGM) Human + ROCKi
3. IntestiCult™ Organoid Differentiation Media (ODM) Human

Select whether the following statement is true or false:

Air-liquid interface (ALI) monolayers are monolayer cultures produced by seeding dissociated intestinal cells onto transwell inserts that provide the option of air-lifting (to expose the apical chamber to the air).

Which of the following statements accurately describes the morphology of healthy, human intestinal organoid cultures:

1. Well defined, smooth epithelium with a central lumen
2. Ruffled membrane with single cells

Which of the following steps for handling reagents during passaging human intestinal organoids is incorrect?

1. Prewetting pipette tips.
2. Thawing Matrigel® on ice.
3. Pre-warming tissue culture treated plates.
4. Using pre-chilled/ cold complete IntestiCult™ media for routine media changes.

Select whether following statement is true or false:

The organoid suspension protocol can be used to set up large numbers of monolayer wells or to generate more organoids for downstream applications.

Which of the following images are not representative of the human intestinal organoid morphology?

1. 
2. 
3. 
4. All of the above are representative culture morphologies.