A refined PCLS extended culture model. (IMAGE)
Caption
(A) A diagram of PCLS generation and key elements of the refined protocol. (B) Donor characteristics of 25 processed human liver tissues and experimental outcomes. (C) Whole plate imaging of PCLS with early culture failure or successful extended culture. (D) Whole-mount dual-colour tissue viability assessment with live cells in green and dead cells in red. (E) Basal OCR of PCLS from healthy or donor with AC measured by Seahorse and corrected for total protein quantity. (F) H&E staining of PCLS over the duration of culture. Arrows point to spaces between hepatocytes seen at day 3 and 5. Dotted lines highlight bridging fibrosis at day 7 and 14. (G) Reticulin staining of both healthy and AC-derived PCLS. The dashed-line highlights a cluster of hepatocytes without basement membrane. (H) KRT19 staining of healthy and AC-derived PCLS. All serial images are from the same donor. *p<0.05; **p<0.01; ***p<0.001. AC, alcohol-associated cirrhosis; AH, alcohol-associated hepatitis; ALD, alcohol-associated liver disease; H&E, haematoxylin and eosin; HBV, hepatitis B virus; KRT19, cytokeratin 19; MASH, metabolic dysfunction associated steatohepatitis; OCR, oxygen consumption rate; PCLS, precision-cut liver slice; PSC, primary sclerosing cholangitis; UW, University of Wisconsin.
Credit
By Z Gordon Jiang, Sofia Izunza Barba, Denisse Rubio-Cruz et al.
Usage Restrictions
Credit must be given to the creator. Only noncommercial uses of the work are permitted.
License
CC BY-NC