Fig. 10.

HA does not directly promote epithelial cell proliferation. Small intestinal epithelial enteroids developed from WT C57BL/6 mice were used to assess the direct effect of HA or PEP-1 on cell proliferation. Enteroid cultures were examined in the stem cell state [50% l-WRN conditioned media (CM)] or in conditions supportive of terminal differentiation (5% l-WRN CM). HA (100 μg/ml) and PEP-1 (125 μg/ml) were added to Matrigel before the enteroids were resuspended. A: enteroid growth as measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) proliferation assay at 72 h in 50% or 5% l-WRN CM. B: RNA expression of Ki-67 and Cyclin-D1 at 72 h expressed as fold change from control enteroids incubated in unsupplemented Matrigel and normalized to GAPDH. Experimental controls include ≥5 wells per condition and were repeated twice. Statistical comparison by unpaired Student's t-test. NS, nonsignificant with P > 0.05.