Cell proliferation and growth curve
Cell growth was measured by MTT [3-(4,5-diethylthiazoly-2-
yl)-2,5-diphenyltetrazolium bromide] assay [14]. Briefly, IECs
(passage 8) were harvested conventionally by digestion and seeded
at 5,000 cells per well in 96-well culture plates in quadruplicate. At
various points in time, medium was removed, and cells were
incubated with 20 ml of MTT solution (5 mg/ml; Sigma) at 37uC
for 4 h, followed by solubilization with 100 ml of 100% dimethyl
sulfoxide (Sigma) at 37uC for 10 min. The absorbance of each well
was measured with a microplate reader (Bio-Tek) at a wavelength
of 570 nm. The viable cell number is proportional to the
absorbance.
Cell proliferation and growth curve
Cell growth was measured by MTT [3-(4,5-diethylthiazoly-2-
yl)-2,5-diphenyltetrazolium bromide] assay [14]. Briefly, IECs
(passage 8) were harvested conventionally by digestion and seeded
at 5,000 cells per well in 96-well culture plates in quadruplicate. At
various points in time, medium was removed, and cells were
incubated with 20 ml of MTT solution (5 mg/ml; Sigma) at 37uC
for 4 h, followed by solubilization with 100 ml of 100% dimethyl
sulfoxide (Sigma) at 37uC for 10 min. The absorbance of each well
was measured with a microplate reader (Bio-Tek) at a wavelength
of 570 nm. The viable cell number is proportional to the
absorbance.
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