Figure 3.
Alginate-Grown HIOs Mature in a Similar Manner as Matrigel-Grown HIOs In Vivo
(A) Dissected kidneys containing tHIOs cultured in alginate or Matrigel. HIOs were cultured in either Matrigel or 2% alginate dissolved in H2O and transplanted after 28 days of culture in vitro. tHIOs were removed after 10 weeks in vivo.
(B) H&E, Alcian blue/PAS, and Trichrome staining reveal the presence of mature crypt-villus structures in alginate and Matrigel tHIOs. Scale bar, 50 μm (H&E and Alcian blue/PAS), Scale bar, 100 μm Trichrome.
(C) Representative images of general epithelial marker staining in tHIOs from alginate and Matrigel. Markers shown are ECAD (epithelial marker), CDX2 (intestinal epithelium marker), PDX1 (duodenum marker), and VIM (mesenchymal marker). Scale bar, 50 μm.
(D) Representative images of specific epithelial cell marker staining in tHIOs from alginate and Matrigel. Markers shown are SOX9 (progenitor cell marker), KI67 (proliferative cell marker), DPP4 (small intestinal brush border enzyme marker), MUC2 (goblet cell marker), CHGA (enteroendocrine cell marker), LYZ and DEFA5 (Paneth cell markers), and OLFM4 (intestinal stem cell marker). Two transplant experiments were conducted with a total of n = 11 Matrigel transplanted and n = 12 alginate transplanted mice. Scale bar, 50 μm for all markers except OLFM4 scale bar, 100 μm.