Modeling human embryo adhesion using a microfluidic platform
For single-cell transcriptomic analysis, stromal and epithelial cells from control and hormone-treated ADOC were collected separately. Stromal cells were first detached by incubation with TrypLE Select® for 15 min and recovered in stromal medium. Subsequently, epithelial cells were dissociated by a 30 min incubation with TrypLE Select® and recovered in organoid medium. Both cell fractions were centrifuged at 300 × g for 5 min at room temperature, the supernatant discarded, and the pellet resuspended in PBS supplemented with 0.04% BSA. Cell concentration and viability were determined using a LUNA™-FL dual fluorescence cell counter (Logos Biosystems) with acridine orange/ propidium Iodide staining (Logos Biosystems, F23001).   Approximately 17,000 cells (combined stromal and epithelial fractions) were loaded onto 10X Chromium G Chip (10X Genomics, PN-10000204) to generate Gel Bead-in-Emulsions (GEMs). Barcoded cDNA libraries were prepared following the manufacturer's instructions using the Chromium Single Cell 3′ Reagent Kit v3.1 (10X Genomics, PN-1000268). Amplified cDNA was quantified with the TapeStation D5000 (Agilent, 5067-5589). Gene expression libraries were constructed with 100-300 ng of amplified cDNA, and library size and concentration were assessed using the TapeStation D5000 kit.