. 2022 Sep;192(9):1200–1217. doi: 10.1016/j.ajpath.2022.05.007

Table 2.

Human Primary Cholangiocarcinoma Cell Lines Established between 2001 and 2021

Cell no.	Year	Country	Cell name	Diagnosis	Isolation source	Digestion method	Sorting method	Maintenance medium	Cell morphology	Doubling time	References
1	2001	Republic of Korea	Choi-CK	IHCCA	Primary tumor	No digestion	Differential trypsinization, scraping, or G418 treatment (100 μg/mL)	Opti-MEM + 10% FBS + 30 mmol/L sodium bicarbonate + antibiotics	Polygonal and compact cells with indistinct cell boundaries	—	75
2			Cho-CK						Polygonal and compact cells with indistinct cell boundaries
3			JCK						Polygonal morphology and some spindle-shaped cells
4			SCK						Monolayer, epithelioid or spindle-shaped cells; fewer polygonal shaped cells
5	2002	Thailand	HubCCA-1	IHCCA	Intrahepatic biliary fluid	—	—	Ham's F12 + 20% FBS + P/S	Adherent, monolayer, polygonal to spindle-shaped cells	—	76,77
6	2002	Thailand	KKU-100	IHCCA	Primary tumor	0.025% trypsin-EDTA, 37°C, 1 hour	Fibroblasts removed with a cell scraper and by differential trypsinization	Ham's F12 + 20% FBS + P/S	Compact, polygonal to spindle-shaped cells; floating or clumping in a confluent monolayer; large nucleus containing two to five nucleoli and a clear cytoplasm	72 hours	77,78
7	2002	Republic of Korea	SNU-245	EHCCA	Primary tumor	No digestion	Differential trypsinization used to remove fibroblasts	RPMI 1640 + 10% FBS	Monolayer, adherent cells; trabecular arrangement with acinar formation	54 hours	79
8			SNU-1079	IHCCA					Monolayer, adherent cells; pleomorphic appearance with multiple cytoplasmic processes; numerous cytoplasmic vacuoles in some cells; some multinucleated cells	72 hours
9			SNU-1196	EHCCA					Monolayer, adherent cells; trabecular pattern consisted of spindle to polygonal shaped cells having vesicular nuclei and multiple small nucleoli	48 hours
10	2004	Japan	TKKK	IHCCA	—	—	—	DMEM (low glucose) + 10% FBS	Adherent, monolayer, epithelial-like cells; pavement-like proliferation	—	80
11	2005	Thailand	KKU-M055	IHCCA	—	—	—	Ham's F-12 + 10% FBS + P/S	—	—	81
12	2005	Thailand	KKU-OCA17	IHCCA	—	—	—	Ham's F-12 + 10% FBS + P/S	—	—	81
13	2005	Japan	TBCN6	EHCCA	Xenograft	0.25% trypsin in PBS, 37°C, 10 minutes	—	DMEM + 10% FBS	Adherent, monolayer, polygonal cells	38 hours	82
14	2005	Japan	TGBC-47	EHCCA	Xenograft	0.25% trypsin in PBS, 37°C, 10 minutes	—	DMEM + 10% FBS	Adherent, monolayer, polygonal cells	38 hours	82
15	2006	Thailand	KKU-M139	CCA	Primary tumor	—	—	Ham's F12 or RPMI 1640 + 10% FBS + P/S	Adherent, monolayer, polygonal cells; pavement-like proliferation	17 hours	16,76
16	2006	Thailand	RMCCA-1	IHCCA	Primary tumor	No digestion	Differential trypsinization used to remove fibroblasts	Ham's F12 + 20% FBS + EGF + 250 μg/mL amphotericin + P/S	Circular to spindle shape with many processes and ornamental fringes; granulated nucleus and cytoplasm	48 hours	83
17	2007	China	HKGZ-CC	IHCCA	Primary tumor	1200–2000 U/mL collagenase and DNase in 10 mL/g DPBS	—	DMEM	Adherent, monolayer, epithelial-like cells	48 hours	84
18	2007	Japan	IHGGK	IHCCA	—	—	—	RPMI 1640 + 10% FBS + P/S	Adherent, monolayer, epithelial-like cells		85, http://www2.idac.tohoku.ac.jp/dep/ccr/TKGdate/TKGvo106/0623.html, last accessed May 5, 2022
19	2009	Thailand	CL-2	CCA	—	—	—	DMEM + 15% FBS + P/S	—	—	86,87
20			CL-6
21			CL-19
22	2010	Japan	NCC-BD1	EHCCA	Xenograft	No digestion	Fibroblasts removed by mechanical scraping	RPMI 1640 + 10% FBS + 2 mmol/L l-glutamine + P/S	Adherent, monolayer, epithelial-like cells; pavement-like proliferation	—	88
23			NCC-BD2	EHCCA
24			NCC-CC1	IHCCA
25			NCC-CC3-1	IHCCA
26			NCC-CC3-2	IHCCA
27			NCC-CC4-1	IHCCA
28	2013	China	HCCC-9810	IHCCA	—	—	—	RPMI 1640 + 10% FBS + P/S	—	20 hours	89
29	2015	Italy	MT-CHC01	IHCCA	Xenograft	200 U/mL collagenase, 3 hours, 37°C	—	Knockout/DMEM/F-12 + 10% FBS + P/S	Monolayer, adherent cells	40 hours	90
30	2016	USA	ICC1	IHCCA	—	Trypsin, 30 minutes, 37°C	—	RPMI or DMEM/F12 + 5% FBS + P/S	—		91
31			ICC2
32			ICC5
33	2016	China	ZJU-0826	EHCCA	Primary tumor	—	Fibroblasts eliminated by differential trypsinization and differential attachment	RPMI 1640 + 10% FBS + antibiotics	Monolayer, adherent, homogeneous cells with characteristic loose pleomorphic cells and rare multinucleated cells	63 hours	92
34	2016	China	ZJU-1125	IHCCA	Primary tumor	—		RPMI 1640 + 10% FBS + antibiotics	Monolayer, epithelial-like, adherent cells; polygonal, occasionally multinucleated	44 hours	92
35	2017	Germany	CCC-5	EHCCA	Pleural effusion	No digestion	—	DMEM + 20% FBS: KFSM - 2:1	Monolayer, spindle- to polygonal shaped cells, pavement-like proliferation; anaplastic, multinucleated giant cells	60 hours	93
36	2017	Thailand	KKU-023	IHCCA	Primary tumor	1 mg/mL collagenase, 37°C, 30–45 minutes	Fibroblasts aseptically removed with a cell scraper	Ham's F12 + 10% FBS + P/S + nonessential amino acids + 12.5 mmol/L HEPES + 50 μg/mL cefazolin + 10 μg/mL ciprofloxacin + 2.5 μg/mL amphotericin B + 5 μmol/L ROCK inhibitor	Ovoid to cuboid shape polygonal cells; seldom multinucleated; forming compact monolayer with occasional multinucleated cells	34 hours	94
37	2017	Thailand	KKU-452	EHCCA	Primary tumor	1 mg/mL collagenase, 37°C, 30–45 minutes	Fibroblasts aseptically removed with a cell scraper		Spindle-shaped cells; seldom multinucleated; segregated and spread surround	17 hours	94
38	2018	China	ICC-1	IHCCA	Primary tumor	No digestion	—	—	—	—	73
39	2018	China	ICC-2	IHCCA	Primary tumor	No digestion	—	—	—	—	73
40	2019	Thailand	KKK-D049	IHCCA	Xenograft	1000 U/mL collagenase + 0.1 mg/mL DNase I, 3 hours	Stromal cells sequentially removed by partial trypsinization and mechanical removal	DMEM + 10% FBS + P/S	Epithelial-like cells; high nuclear to cytoplasmic ratio; tight clustering	—	95
41			KKK-D068						Epithelial-like, polygonal and spindle-shaped cells; high nuclear to cytoplasmic ratio	—
42			KKK-D131						Epithelial-like, polygonal and spindle-shaped cells; high nuclear to cytoplasmic ratio	—
43			KKK-D138						Epithelial-like, polygonal and spindle-shaped cells; high nucleus-to-cytoplasm ratio	—
44	2021	Italy	82.3	IHCCA	Xenograft	Collagenase 200 U/mL, 3 hours, 37°C		10% FBS + P/S + gemcitabine	Monolayer, adherent, epithelial-like cells	53 hours	74

CCA, cholangiocarcinoma; DMEM, Dulbecco's modified Eagle's medium; DPBS, Dulbecco's phosphate-buffered saline; EGF, epidermal growth factor; EHCCA, extrahepatic cholangiocarcinoma; FBS, fetal bovine serum; IHCCA, intrahepatic cholangiocarcinoma; KFSM, keratinocyte serum free medium; MEM, minimal essential medium; PBS, phosphate-buffered saline; P/S, penicillin/streptomycin; ROCK, Rho-associated kinase.