Fig. 2.

50% L-WRN CM is required for LF/SC enteroid growth and expansion. (A) Enteroids from LF/SC animals cultured with 50% L-WRN CM expand soon after crypt isolation (Day 1), growing to large size after passage (Day 6), and continue to expand and obtain crypt-like morphology at higher passage (white arrows) (P5 and P>10). Lower magnification shows enteroids were maintained at high density throughout passage history (P>10, 500 µm). Enteroids fail to grow when cultured in D10 after isolation (Day 6) or when passed to 50% L CM after high passage (P>10) growth in 50% L-WRN CM, no data (N/D). (B) Fold expansion of wells containing LF/SC enteroids. LF/SC enteroids grown in 24-well plates were maintained at high density. The ratio of per well expansion was recorded at each passage throughout the passage history. (C) Number of enteroids expands with each passage. A total of 200 high passage (P>10) enteroids were seeded to two wells and the number of enteroids was counted after 3-4 days of growth, and again after subsequent passage splitting 1:2, n=3. Cat samples are not included in B or C due to the lower passage and density of enteroids. Error bars indicate ±s.d.