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.