eCHIP Culture System and its Response to Reproductive Hormones

First, the eCHIP culture system was established as described previously, and parameters were set to control the flow rate between 1-10 ul/min in a sinusoidal pattern.

After 3 days of growth in ExM/ECM media, the inlet reservoir was changed to different media for two groups. The control group received ExM/ECM mixed media, while the estrogen group received ExM/ECM mixed media containing 10nM E2.

After 48 hours, the medium of the estrogen group was changed to differentiation medium (DM), which consisted of ExM/ECM mixed media containing 10nM E2, 1uM P4, 1uM cAMP, and 20ng/ml PRL. This group was referred to as the decidual group and was cultured for an additional 4 days.

For the detection of PRL in supernatants, an ELISA method was used. The differentiation protocol for the decidual group was modified to include ExM/ECM mixed media containing 1uM E2, 1uM MPA, and 0.5mM cAMP, based on a previously reported protocol2.

The response of eCHIP to reproductive hormones will be examined subsequently using IF, IHC, PAS, and ELISA experiments.