Establishment of various biliary tract carcinoma cell lines and xenograft models for appropriate preclinical studies

Table 1 Clinicopathological features of original biliary tract tumors

Xenograft	Pathological diagnosis of original tumor	Age/sex	Histologic type	Prognosis (survival days)	Chemotherapy	Clinical evaluation of chemotherapy effect (effective days)	Established cell line
1	CCC	70/F	Adeno, mod	Death (402)	Non		NCC-CC1
2	CCC	71/F	Adeno, mod	Death (175)	Non		NCC-CC3-1
							NCC-CC3-2
3	CCC	59/M	Adeno, mod	Alive (2172)	Non		NCC-CC4-1
							NCC-CC4-2
							NCC-CC4-3(NCC-CC5)
4	CCC	31/M	Adeno, mod + PSC	Death (386)	GEM + TS1	SD (84 d)	NCC-CC6-1
							NCC-CC6-2
5	Distal BDCa	58/F	Adeno, mod	Death (299)	GEM	PD	NCC-BD1
6	Distal BDCa	77/F	Adeno, mod	Death (393)	GEM	PD	NCC-BD21
7	Distal BDCa	80/M	Adeno, mod	Death (212)	Non		NCC-BD3
8	Hilar BDCa	74/M	Adeno, mod	Death (172)	Non		NCC-BD4-1
							NCC-BD4-2
9	Hilar BDCa	48/M	Adeno, well	Alive (500)	GEM	PD	NA
10	Hilar BDCa	43/M	Adeno, mod	Alive (1422)	Non		NA
11	CCC	69/M	Adeno, mod	Death (174)	Non		NA
12	CCC	54/F	Adeno, mod	Death (181)	Non		NA
13	CCC	56/M	Adeno, mod	Death (319)	GEM	PD	NA
14	CCC	73/M	Adeno, mod	Death (53)	Non		NA
15	CCC	54/M	Adeno, mod	Alive (2608)	Non		NA
16	CCC	45/F	Adeno, mod	Alive (882)	GEM + CDDP	Unknown	NA
17	CCC	72/M	Muc	Death (749)	GEM/GEM + TS1	Unknown	NA
18	CCC	78/M	Adeno, mod	Death (382)	GEM	Unknown	NA
19	CCC	66/M	Adeno, mod	Death (168)	Non		NA
20	CCC	65/M	CoCC	Alive (1604)	Non		NA
21	CCC	70/M	Adeno, por	Death (851)	GEM	SD (49 d)	NA
22	CCC	63/F	Adeno, mod	Alive (363)	Unknown	Unknown	NA
23	CCC	72/M	Adeno, mod	Death (394)	GEM	PD	NA
24	CCC	77/F	Adeno, mod	Death (445)	GEM	SD (105 d)	NA
25	Hilar BDCa	66/M	Adeno, mod	Alive (102)	GEM + TS1	Unknown	NA
26	Distal BDCa	54/M	Adeno, mod	Alive (2096)	Non		NA
27	Distal BDCa	67/M	Adeno, mod	Death (672)	GEM + TS1	PD	NA
28	Distal BDCa	80/M	Adeno, mod	Alive (2024)	GEM	PR-CR (548 d)	NA

¹BD2 was obtained from the direct culture of patient specimens. CCC: Cholangiocellular carcinoma; BDCa: Bile duct carcinoma; Adeno: Adenocarcinoma; mod: Moderately differentiated; PSC: Primary sclerosing cholangitis; Muc: Mucinous carcinoma; CoCC: Cholangiolocellular carcinoma; por: Poorly differentiated; non: No chemotherapy received; GEM: Gemcitabine; CDDP: Cisplatin; SD: Stable disease; PD: Progressive disease; PR: Partial response; CR: Complete response.

Table 2 Sensitivity to gemcitabine in each cell line

Cell line	Sensitivity to gemcitabine in cell line1
	IC50 (μmol/L)	IC60 (μmol/L)	IC70 (μmol/L)	IC80 (μmol/L)
NCC-CC1	86.78	N.A	N.A	N.A
NCC-CC3-1	0.04	1.82	9.31	85.21
NCC-CC3-2	0.10	1.92	43.83	N.A
NCC-CC4-1	0.05	4.08	N.A	N.A
NCC-CC4-2	0.03	11.53	N.A	N.A
NCC-CC4-3 (NCC-CC5)	0.06	4.92	95.10	N.A
NCC-CC6-1	0.01	0.02	0.06	3.76
NCC-CC6-2	10.98	35.67	N.A	N.A
NCC-BD1	7.66	58.00	N.A	N.A
NCC-BD2	N.A	N.A	N.A	N.A
NCC-BD3	N.A	N.A	N.A	N.A
NCC-BD4-1	0.04	0.06	0.09	2.93
NCC-BD4-2	0.06	0.07	0.19	5.37

¹The cytotoxicity of gemcitabine for each cell line was assessed by a modified 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt assay with CellTiter 96 Aqueous One Solution Reagent (Promega, Madison, WI, United States). Tumor cells (3000 cells⁄well) in the exponential growth phase were grown in 96-well plates. IC: Inhibitory concentration.