Figure 3.

Comparison of cell viability between whole tissue versus tissue dissected into smaller pieces

(A) Nanosight (NTA) profiles and protein concentration by bicinchoninic acid (BCA) assay for human colon that was cultured as whole tissue or dissected into smaller pieces.

(B) Immunoblotting of lysates (20 μg of proteins) from baseline tissue (directly after collection, timepoint 0 h) immediately placed into lysing buffer without culturing, whole tissue, and dissected tissue (after 12 h culture) derived from the human colon. PARP antibody (#9542, Cell Signaling Technology [CST]), cleaved caspase-3 antibody (#9661S, CST), GAPDH antibody (#2118S, CST) were used for immunoblotting.

(C) Comparison of human colon whole tissue and dissected tissue cell viability as determined by dual Acridine Orange / Propidium Iodide fluorescence and brightfield optics, using the LUNA automated cell counter. Live cells are green and dead cells are red.

(D) Comparison of human colon whole tissue and dissected tissue cell viability by apoptotic flow staining shows similar viability between baseline tissue directly after collection and whole and dissected tissues after 12 h tissue explant culture.

(E) Comparison of cell viability between baseline and 24 h cultures of mouse lung tissue explant.

(F) Cell viability, as determined by flow cytometry with LIVE/DEAD™ Fixable Blue Dead Cell Stain, shows increased viability in dissected tissue cultures compared to whole tissue culture from mice.