Development of Collagen-Based 3D Matrix for Gastrointestinal Tract-Derived Organoid Culture

Figure 1

Effect of the components of collagen-based matrix on culturing mouse colon organoids. (a) Comparison of Matrigel (upper panel), collagen-based matrix without Ham’s F12 (middle panel), and collagen-based matrix with Ham’s F12 (down panel) shows different phenotypes. All matrices were mixed with the organoid at a ratio of 1 : 1. (b) Diverse percentages of collagen (30%, 70%, and 80%) distinguish different growth in each percentage. Organoids in Passage 0 were followed up on under a microscope daily. (c) Measured circumstance of the organoids shows the effect of the percentage of Ham’s F12 (0%, 5%, and 10%). (d) Measured circumstance of the organoids with each collagen percentage rate (30%, 70%, and 80%). (e) Concentration of bicarbonate was tested and followed up daily under a microscope. (f) Measured circumstances of the organoids in different concentrations of bicarbonate were similar. (g) Relative mRNA expression in representative markers of the organoids was measured in Matrigel with 0%, 5%, and 10% bicarbonate. Lgr5 was used as a marker of intestinal stem cells, E-cadherin (Ecad) as a marker of epithelial cells, villin-1 (Vil1) as a marker of the epithelium border, defensin-5 (Defensin5) as a marker of Paneth cells, and chromogranin A (ChrgA) as a marker of enteroendocrine cells. The circumstance of the organoids was measured on the last day in vitro. Day denotes days of in vitro. Bar scale is 50 μm. , , .

(a)

(b)

(c)

(d)

(e)

(f)

(g)