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After centrifugation (450 x g, 30 min, and 20 C), the white blood interface was washed in PBS 1x (200 x g, 15 min, and 20 C) and the cell pellet was resuspended in IMDM medium supplemented with 10% FBS and 50 g/mL gentamicin to a concentration of 500,000 cells/mL

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Supporting this view, many anterograde tracing and immunohistochemistry studies, with the exception of two studies 38,39 , have consistently shown that axons in the intestinal mucosa closely approach enteroendocrine cells without establishing contacts 22,24,27,40,41,42,43,44