Establishment of a highly differentiated immortalized human cholangiocyte cell line with SV40T and hTERT

Maruyama, Masanobu; Kobayashi, Naoya; Westerman, Karen A.; Sakaguchi, Masakiyo; Allain, Jean E.; Totsugawa, Toshinori; Okitsu, Teru; Fukazawa, Takuya; Weber, Anne; Stolz, Donna B.; Leboulch, Philippe; Tanaka, Noriaki

doi:10.1097/01.tp.0000110292.73873.25

Cited by 122 publications

(90 citation statements)

References 32 publications

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“…We devised a strategy for differentiation of ES cells into hepatocyte-like cells using a four-step differentiation protocol: (i) generation of EBs; (ii) induction of definitive endoderm from 2-d-old EBs by growth in Activin A and FGF-2; (iii) induction of hepatic progenitors by co-culture with human liver nonparenchymal cell lines (human liver endothelial (TMNK-1) 14 , stellate (TWNT-1) 15 and cholangiocyte (MMNK-1) 16 cell lines) and a deleted variant of HGF (dHGF); and (iv) maturation into hepaticlike cells by culture in DMSO and dexamethasone (Fig. 1).…”

Section: Protocols Have Been Developed To Differentiate and Enrich Vamentioning

confidence: 99%

“…Induction of hepatic progenitor cells (expressing both albumin and AFP) and maturation into hepatocyte-like cells by co-culture with human liver nonparenchymal cell lines 12| Prepare human liver nonparenchymal cell lines [14][15][16] , which can be maintained in T75 tissue culture flasks in 10 ml of DMEM and 10% FBS. Passage confluent cultures every week, and change the medium every 3 d.…”

Section: |mentioning

confidence: 99%

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Differentiation of mouse embryonic stem cells to hepatocyte-like cells by co-culture with human liver nonparenchymal cell lines

et al. 2007

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This protocol describes a co-culture system for the in vitro differentiation of mouse embryonic stem cells into hepatocyte-like cells. Differentiation involves four steps: (i) formation of embryoid bodies (EB), (ii) induction of definitive endoderm from 2-d-old EBs, (iii) induction of hepatic progenitor cells and (iv) maturation into hepatocyte-like cells. Differentiation is completed by 16 d of culture. EBs are formed, and cells can be induced to differentiate into definitive endoderm by culture in Activin A and fibroblast growth factor 2 (FGF-2). Hepatic differentiation and maturation of cells is accomplished by withdrawal of Activin A and FGF-2 and by exposure to liver nonparenchymal cell-derived growth factors, a deleted variant of hepatocyte growth factor (dHGF) and dexamethasone. Approximately 70% of differentiated embryonic stem (ES) cells express albumin and can be recovered by albumin promoter-based cell sorting. The sorted cells produce albumin in culture and metabolize ammonia, lidocaine and diazepam at approximately two-thirds the rate of primary mouse hepatocytes.

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Section: Protocols Have Been Developed To Differentiate and Enrich Vamentioning

confidence: 99%

Section: |mentioning

confidence: 99%

Differentiation of mouse embryonic stem cells to hepatocyte-like cells by co-culture with human liver nonparenchymal cell lines

et al. 2007

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“…The KKU-M213 cell line, which was established from a primary tumor mass of intrahepatic CCA (11), was provided by the Liver Fluke and Cholangiocarcinoma Research Center, Faculty of Medicine, Khon Kaen University (Khon Kaen, Thailand). An immortalized cholangiocyte cell line, MMNK-1, which was established by Maruyama et al (12), was used as the control. The two cell lines were cultured in Ham's F-12 (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) culture medium supplemented with 10% fetal bovine serum (Gibco; Invitrogen; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin and 100 µg/ml streptomycin.…”

Section: Methodsmentioning

confidence: 99%

High expression of CCDC25 in cholangiocarcinoma tissue samples

et al. 2017

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Abstract. Cholangiocarcinoma (CCA) is a malignant transformation of biliary epithelial cells. It is a slow growing tumor, but is also highly metastatic with a poor prognosis. Bile acids are known to transactivate the epidermal growth factor receptor (EGFR) in cholangiocytes and induce cyclooxygenase-2 expression. The protein expression profiles of bile acid-treated CCA cells were studied using a proteomic approach. To elucidate the possible mechanisms involved in the bile acid-mediated enhancement of CCA cell migration, the effects of six bile acids, including cholic, deoxycholic, taurocholic, taurodeoxycholic, glycocholic and glycodeoxycholic acid, on the migration of CCA cells were examined in vitro using wound healing assays. Subsequently, the possible proteins involved in enhanced CCA cell migration were investigated using a proteomic approach. Changes to the protein expression profiles of CCA cells following bile acid treatment was examined using two-dimensional electrophoresis and mass spectrometry. The results demonstrated that cholic and deoxycholic acid significantly enhanced the migration of CCA cells, compared with the treated MMNK-1 control cells. CCA cells had 77 overexpressed protein spots following cholic acid treatment, and 50 protein spots following deoxycholic acid treatment, compared with the treated MMNK-1 control cells. Liquid chromatography tandem-mass spectrometry analysis revealed that coiled-coil domain containing 25 (CCDC25) was significantly overexpressed in cholic acid-treated CCA cells compared with in cholic acid-treated control cells. When the expression levels of CCDC25 were investigated using western blot analysis, CCDC25 was demonstrated to be highly expressed in CCA tissues, but not in the adjacent non-cancerous tissue samples. The identified proteins were further analyzed for protein-chemical interactions using STITCH version 3.1 software. CCDC25 protein was identified to be associated with Son of sevenless homolog 1 and growth factor receptor-bound protein 2, which are involved in EGFR signaling. The results of the present study demonstrated that following cholic acid treatment, CCDC25 is overexpressed in CCA cells, which is associated with significantly enhanced cell migration. This suggests that CCDC25 is a potential therapeutic target for the treatment of patients with CCA. IntroductionCholangiocarcinoma (CCA) is a malignant transformation of biliary epithelial cells. It is a slow growing tumor, but it is also highly metastatic with a poor prognosis (1). The highest incidence of this cancer has been identified in Asia, particularly in Northeast Thailand (2). In the majority of cases of CCA, the etiology is unknown. Chronic inflammation and cellular injury within bile ducts, together with the partial obstruction of bile flow result in high-risk conditions for CCA development (3). The tumor not only may develop mechanisms to survive in the toxic environment of bile, but may also utilize bile components to promote cell growth, survival and invasion (4-6). Bile acids are known...

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“…of Surgery, Okayama, Japan) 38 . Primary human bile duct epithelial cells were donated by Dr. H. Crosby, Univ.…”

Section: In Vitro Experimentsmentioning

confidence: 99%

Inhibition of Integrin αvβ6 on Cholangiocytes Blocks Transforming Growth Factor-β Activation and Retards Biliary Fibrosis Progression

et al. 2008

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Establishment of a highly differentiated immortalized human cholangiocyte cell line with SV40T and hTERT

Abstract: We have established an immortalized cholangiocyte cell line, MMNK-1, using SV40T and hTERT transduction.

Cited by 122 publications

References 32 publications

Differentiation of mouse embryonic stem cells to hepatocyte-like cells by co-culture with human liver nonparenchymal cell lines

Differentiation of mouse embryonic stem cells to hepatocyte-like cells by co-culture with human liver nonparenchymal cell lines

High expression of CCDC25 in cholangiocarcinoma tissue samples

Inhibition of Integrin αvβ6 on Cholangiocytes Blocks Transforming Growth Factor-β Activation and Retards Biliary Fibrosis Progression

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