In vivo development of immune tissue in human intestinal organoids transplanted into humanized mice
O), for 5 min at room temperature. An isovolume of PBS was added and tubes were centrifuged at 300for 5 min. The pellet was washed in PBS, centrifuged and resuspended in FACS buffer .
After digestion of the tissue at 37 °C for 30 min with DNAse I and Collagenase A , the homogenate was filtered and centrifuged at 450Both HIO and SI cell suspensions were individually resuspended in 44% Percoll , loaded on a 67% Percoll layer and centrifuged at 650without brake for 30 min at room temperature. The cell layer at the interface of the two gradients was collected, washed, counted and used for in vitro assays, or stained for immune markers and analyzed by mass cytometry.
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