Adult human liver slice cultures: Modelling of liver fibrosis and evaluation of new anti-fibrotic drugs

Table 1 METAVIR scores and description of clinical liver samples

METAVIR score		Patients (n)	Pathology
F0-F1	No fibrosis or mild fibrosis	10	HBV-, HCV-, HIV-seronegative patients who underwent liver resection surgery, mainly for liver metastasis, in the absence of underlying liver disease. A0-F1
F0-F1	No fibrosis or mild fibrosis	1	Prior history: Breast cancer with liver metastases, treated by surgery and radio-chemotherapy. Non-tumoral liver sample: Perisinusoidal and portal fibrosis without septa (F1). No steatosis
		2	Prior history: HCV infection, resected hepato-cholangiocarcinoma. Non tumoral liver samples: A0F0
F2-F3	Moderate to severe fibrosis	2	Cholangiocarcinoma, non-tumor liver samples
		1	Chronic hepatitis B infection, NASH, and two resected hepatocellular carcinoma nodules. Non-tumoral liver sample: Chronic hepatitis with extensive fibrosis A1F3
F4	Cirrhosis	2	HCC, non-tumor liver samples
		1	HCC, treated HCV infection. Non-tumoral liver sample
		1	HCC on untreated HCV infection. Non-tumoral liver sample

A “significant” fibrosis, as defined by a fibrosis grade (F), is greater than 1 by the METAVIR scoring system, with usually a significant necrotic inflammation as defined by an activity grade (A) greater than 1 by the METAVIR scoring system. Fibrosis grade: F0: No fibrosis, F1: Portal fibrosis without septa, F2: Portal fibrosis with few septa, F3: Numerous septa without cirrhosis, F4: Cirrhosis. Activity grade: A0: No activity, A1: Mild activity, A2: Moderate activity, A3: Severe activity. HBV: Hepatitis B virus; HCV: Hepatitis C virus; HCC: Hepatocellular carcinoma; HIV: Human immunodeficiency virus; NASH: Nonalcoholic steatohepatitis.

Table 2 Primers used for real-time reverse transcription-quantitative polymerase chain reaction analysis

Gene	Forward primer sequence	Reverse primer sequence
CYP2E1	AGCACAAACTCTGAGATATGG	ATAGTCACTGTACTTGAACT
CYP3A4	GCCTGGTGCTCCTCTATCTA	ACAGGCTGTTGACCATCATAAAAG
HNF-1β	ACGTCAGAAAGCAACGAGAGATC	CCCAGGCCCATGGCT
HNF-4α	CCTGGAATTTGAGAATGTGCAG	AGGTTGGTGCCTTCTGATGG
Albumin	ATGAGATGCCTGCTGACTTG	GCACGACAGAGTAATCAGGA
18S RNA	CAGAGCGAAAGCATTTGCCAAG	CGGCATCGTTTATGGTCGGAAC
TGF-β1	CCTGGAAAGGGCTCAACAC	CAGTTCTTCTCTGTGGAGCTGA
HSP 47	GCCACCGTGGTGCCGCA	GCCAGGGCCGCCTCCAGGAG
β-SMA	AGGGGGTGATGGGTGGGAA	ATGATGCCATGTTCTATCGG
Procol1A1	CAATCACCTGCGTACAGAACGCC	CGGCAGGGCTCGGGTTTC
MMP-2	CTT CGCCCC AGG CAC TGG TG	CCTCGCTCCCATGGG GTT CGGT
MMP-9	GGT CCCCCCACT GCT GGC CCTTCTACGGCC	GTCCTCAGG GCACTG GAG GAT GTC ATA GCT
VEGF	TACCTCCACCATGCCAAGTG	ATGATTCTGCCCTCCTCCTTC
GAPDH	ACCAGGGCTGCTTTTAACTCT	GGTGCCATGGAATTTGCC