Low-level, interventional, biological and non-pharmacological study prospective intervention for the preparation of organotypic human liver slice cultures Liver Slice Culture (hLSC) applying the protocol described by Jiang and collaborators.
Human liver slice culture hLSCs will be used from non-tumor liver tissue collected by biopsy from patients undergoing abdominal surgery, who have signed informed consent and agree to perform a liver biopsy for the specific research protocol. The study is "low level interventional" when a small portion (approximately 10 mm in diameter) of liver tissue macroscopically free from parenchymal alterations obtained in the open at the site of surgery for primary or secondary liver tumors will be taken (hepatic metastases from colorectal, pancreatic or breast cancer). The goal is to create organotypic liver cultures (hLSC) derived from liver biopsies taken from patients undergoing abdominal surgery. for primary or secondary liver tumors for the molecular characterization of liver fibrosis. Use hLSC to study the liver microenvironment and molecular determinants involved in fibrosis through single cell sequencing; compare the sequencing data obtained with hLSC with that obtained via characterization of a GEMM whose generation is foreseen in the same project financed by the MoS. To generate this GEMM, the pro-fibrotic Hipk2 gene was specifically deleted in the tissue liver (HIPK2-liver knock-out, HIPK2-LKO) and liver fibrosis will be induced through a high-fat, low-choline diet.
Study Type
INTERVENTIONAL
Allocation
NA
Purpose
DIAGNOSTIC
Masking
NONE
Enrollment
80
A small portion (about 10 mm in diameter) of liver tissue was collected macroscopically free from alterations of the parenchyma obtained in the open at the site surgery for primary or secondary liver tumors, for the preparation of cultures of organotypic liver slices.
"Regina Elena" National Cancer Institute
Rome, Italy
RECRUITINGMolecular characterization of liver fibrosis
Creation of organotypic cultures of liverderived from liver biopsies taken from patients undergoing abdominal surgery for primary or secondary liver tumors for the molecular characterization of liver fibrosis.Patients will undergo abdominal surgery (as per clinical practice), during which a biopsy will be performed in an area of the liver macroscopically free from parenchymal alterations, approximately 20/25 slices will be obtained from each sample. The hLSCs will come used to study the hepatic microenvironment in the presence or absence of pro-fibrinogenetic stimuli e of anti-fibrotics by time-lapse analysis and confocal immunofluorescence microscopy multiple and through single cell sequencing.
Time frame: 24 months
Study of the hepatic microenvironment
Use hLSCs to study the liver microenvironment and the molecular determinants involved in it fibrosis through single cell sequencing (single-cell RNA sequencing);
Time frame: 24 months
Compare the sequencing data
Compare the sequencing data obtained with hLSCs with those obtained through the characterization of a GEMM. A biopsy will be performed after abdominal surgery (as per clinical practice), in an area of the liver macroscopically free from parenchymal alterations. hLSCs will be analyzed over the following week for viability and fibrosis induction using fibrosis-specific stains for time-lapse dynamic imaging (IncuCyte), i.e. picrosirius red, Masson's trichrome, and quantitative RT-PCR and multiplex immunofluorescence
Time frame: 24 months
This platform is for informational purposes only and does not constitute medical advice. Always consult a qualified healthcare professional.