Figure 2.
(A) Example of crypt isolated using digestive solution. Arrow (red) shows crypts obtained from the duodenal tissue. (B) Images of enteroids obtained from fresh tissue (Angus steer) and cryopreserved tissue (Holstein calf) from d 1 to 7 in passages 0 and 1. (C) Budding of crypt domains of enteroid on d 5. A white asterisk indicates a budding crypt domain. (D) Confocal images of enteroids (d 3 in passage 0 derived from fresh [left] and cryopreserved [right] tissue). Nuclei were stained by DAPI (blue; Sigma-Aldrich), and F-actin in the brush border microvilli was stained using Texas Red-conjugated phalloidin (red; Thermo Fisher Scientific). Enteroid area (μm2) in each calf and passage from d 1 to 7 from fresh tissues from Angus steers (n = 3 [n = 2 in passage 1]; E) and cryopreserved tissues from Holstein calves (n = 3 [n = 1 at d 7 in passage 1]; F).