Factors affecting in vitro growth of harvested enterocytes

Abstract

Selective enterocyte transplantation may be an alternative to whole organ transplantation for increasing absorptive capacity. Our aim was to determine the effect of initial cell number and viability, proportion of intact crypts, and basement membrane components (BMC) on the in vitro growth of rabbit enterocytes. Enterocytes were harvested using warm trypsinization from ileal segments in 40 rabbits. Initial cell viability was 92 +/- 4% (mean +/- SD), cell yield was 7.7 +/- 3.6 x 10(6) cells/cm, and there were 0.51 +/- 0.33 crypts/100 cells. Initial cell viability correlated with cell yield (r = -0.508, p < 0.001) and % crypts (r = 0.313, p < 0.05). Cell yield also correlated with % crypts (r = -0.645, p < 0.001). Enterocytes (5 x 10(6)) were incubated in growth media in plain or BMC coated growth culture vessels for 14 days. There was a correlation between both number of cells seeded (r = 0.824, p < 0.001) and cell viability (r = -0.696, p < 0.01) and % growth colonies containing epithelial cells at 14 days. Both total growth colonies (r = -0.565, p < 0.05) and colonies with epithelial cells (r = -0.589, p < 0.05) had a negative correlation with % crypts. Incubating cells in BMC coated vessels (n = 6) resulted in significantly more dispase liberated cells after 14 days than in plain vessels (n = 6) (6.6 +/- 1.1 vs. 3.8 +/- 1.0 x 10(6), p < 0.05) but viability was similar (97 +/- 2% vs. 96 +/- 2%).(ABSTRACT TRUNCATED AT 250 WORDS)