Figure 1. Precision Cut Intestinal Slice (PCIS) preparation, viability, and cellular distribution
(A) After surgical excision of a portion of intestinal tissue as part of a clinically indicated procedure, a sample (approximately 1.0 x 0.5 x 0.2cm) was taken from the proximal site of the excision and placed in oxygenated, chilled, buffer (I) . The tissue was dissected into smaller strips (approximately 0.5 x 0.2cm), embedded in low melting temperature agarose (II) and 400µm thick slices(III) were generated. PCIS were kept in an oxygenated incubator(IV) (37oC, 90% O2, 5% CO2). (B) Cell death in PCIS was determined using the lactate dehydrogenase (LDH) assay. Absorbance was measured as OD 490nm (n=16). Detergent Triton X-100 (TX-100) used as a positive control (C) Metabolic activity in PCIS was measured using the water-soluble tetrazolium salt (WST-1) assay. Absorbance was measured as OD 450nm (n=16). (D) Single nuclear RNA sequencing gene expression data of human colon tissue samples (n=4) merged to display approximate cellular distribution of PCIS using Uniform Manifold Approximation and Projection. *P <.05, ****P<.0001, bars indicate mean, error bars represent ± SEM.