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Transwell chambers were assayed for cell invasion ability by applying 100 L of DMEM-diluted Matrigel gel (3:7 dilution) to the chambers to be checked and incubated at 37 C for 30 min to form a gel, then pre-treated MIA PaCa-2 and PANC-1 cells were resuspended with DMEM and spread in small chambers at 50000/well, and the lower chamber was placed in 20% FBS DMEM, incubated at 37C 5% CO 2 for 48 hours

However, as discussed in Change Requests 12970 and 13103, if a MDH cancelled its rural classification under 412.103(g) effective on or after October 1, 2022, its MDH status may not be applied continuously or automatically reinstated, as applicable (and as described previously)

doi: 10.1007/s40520-019-01223-5 34 YanMSuJLiY