In vitro enterocyte-like differentiation and co-culture

Cells were divided into two groups. Briefly, in group A, rBMSCs of passage 3 were inoculated at 1×10⁶ cells/cm² in six-well culture plates in the lower chamber of the Transwell (Costar, Corning, NY, USA). Then, the Transwell insert was placed in the hole, and mIEC-6s of passage 3 were inoculated in the upper chamber. The co-culture system of rBMSCs and mIEC-6s was established by mixing together the upper and lower chambers containing DMEM (HyClone Corp, Logan, Utah, USA). In group B, the control group, rBMSCs of passage 3 were inoculated both in upper and lower chambers in the same way. All groups were cultured at 37°C and 5% CO₂ for 3, 7, or 10 days. They were placed under a fluorescence microscope camera for immunofluorescence detection.