Figure 1.

Immune cell function in cryopreserved human precision-cut lung slices (hPCLSs). (A) Phagocytosis assay in thawed hPCLSs. Left panel shows a representative image of intracellular fluorescence of in situ phagocytes after 90-minute incubation of hPCLSs with pHrodo red–conjugated Escherichia coli particles indicative of active phagocytosis. Representative fluorescence images show the same hPLCS stained with CD11c antibody (middle panel, in green), followed by phagocytosis assay (right panel). A large majority of CD11c⁺ cells had intracellular pHrodo red E. coli particles. (B) Representative images showing (left panel) cells on the culture plate after thawed hPCLSs were cultured for 6 days in control media (RPMI + 10% FBS) or (right panel) under dual activation by IL-2 (10 ng/ml) and anti-CD3 (2 μg/ml) plus anti-CD28 (8 μg/ml) antibodies coated on the bottom of culture plates to stimulate lymphocytes. (C) Proliferation analysis of CD4 gated lymphocytes with carboxyfluorescein succinimidyl ester dilution and flow cytometry. The number of cell division was marked. (D) Flow cytometry analysis of lymphocytes on the plates of control and activated hPCLS cultures. Cells gated with CD4 and CD8 were assayed for their activation status by their expression of CD69 and CD25.