Figure 1. Organoids Grown in IntestiCult™-SF Have a Comparable Morphology to Organoids Grown in IntestiCult Organoid Growth Medium (Human)
Human intestinal organoids grown in IntestiCult™-SF and IntestiCult™ Organoid Growth Medium (Human) display similar morphological characteristics during organoid maintenance and expansion. Shown are organoids from small intestinal and colonic tissues, established and expanded in IntestiCult™-SF or IntestiCult™ OGMH. Organoids are imaged at passage 5 of culture for all images. Scale bars = 500 μm.
Figure 2. IntestiCult™-SF Provides More Efficient Expansion of Intestinal Organoids Compared to IntestiCult™ OGMH
Human intestinal organoids established from both (A) small intestinal and (B) colonic tissues display more efficient expansion during extended culture when grown in IntestiCult™-SF compared to those grown in IntestiCult™ OGMH. Shown is the cumulative expansion of organoids averaged across two separate donor lines in each medium. IntestiCult™-SF demonstrates more efficient expansion of both small intestinal and colonic organoids during extended culture.
Figure 3. Organoids Grown in IntestiCult™-SF Display Some Characteristics of the Mature Intestinal Epithelium
While organoids grown in IntestiCult™-SF display a primarily "immature" phenotype including expression of (A) intestinal stem cell marker OLFM4 and (B) proliferation marker ki67. (C, D) These organoids also display some characteristics of the mature epithelium such as the presence of goblet cells (C, Muc2), enterocytes (C, KRT20), and enteroendocrine cells (D, CHGA). Scale bars = 50 μm or 20 μm as indicated.
Figure 4. Organoids Grown in IntestiCult™-SF Display Similar Genetic Expression Profiles to Those Grown in IntestiCult™ OGMH
Analysis of intestinal organoids grown in IntestiCult™-SF by qPCR of (A) Lgr5, (B) Muc2, (C) Krt20, and (D) ChgA demonstrates similar genetic expression profiles to those grown in IntestiCult™ OGMH, as well as to colonic and small intestinal tissue (A-D; x and + respectively). All expression levels are shown relative to ACTB and TBP house-keeping genes (HKGs). (E) Further analysis of three separate donors grown in IntestiCult™-SF (circles) IntestiCult™ OGMH (triangles), and published medium (squares) by principle component analysis demonstrates that differences between cultures are primarily due to donor variability with samples forming distinct clusters separated by donor, rather than by medium.
Figure 5. IntestiCult™-SF Enables Efficient Expansion of Intestinal Organoids from Polyp Tissue
Intestinal organoids were established from both normal epithelial tissue, as well as from polyp tissue, and expanded for three passages in IntestiCult™-SF. Shown are cultures from two separate donors, imaged at the end of each passage immediately before passaging.
Figure 6. Organoids Grown in IntestiCult™-SF can be Further Differentiated using IntestiCult™ Organoid Differentiation Medium
Further differentiation of organoids in IntestiCult™-SF can be achieved by passaging organoid cultures in IntestiCult™ Organoid Differentiation Medium as organoid-derived monolayers (2D Monolayer Diff) or in 3D organoid culture (3D Organoid Diff). Upon differentiation, markers for enterocytes (KRT20, ApoB), goblet cells (Muc2), and enteroendocrine cells (ChgA) are upregulated compared to organoids grown in IntestiCult™-SF.
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