Atcc highly recommends that protective gloves and clothing always be used and a full face mask. Hep2 atcc ccl23 homo sapiens hela contaminant carcinoma. Hep g2 or hepg2 is a human liver cancer cell line hep g2 is an immortal cell line which was derived from the liver tissue of a 15yearold african american adolescent boy with a welldifferentiated hepatocellular carcinoma. Hepatocellular carcinoma hcc is the most common liver cancer and has a poor prognosis. Atcc confirmed this cell line is positive for the presence of human papilloma viral dna sequences via pcr.
Data on the expression of pepck in hepg2 hepatocytes. C3a hepg2 c3a, derivative of hep g2 atcc hb8065 atcc crl10741 frozen forprofit. Forkhead box a1 foxa1 expression is associated with various types of tumors. So, if you need liver cells, hep2 are a bad choice. Example protocol for the culture of the hepg2 cell line on alvetex 22 mm disc in 6well insert format, ams.
The parental cells were stably transfected at passage 48 with a human cholesterol 7 alphahydroxylase cyp7 minigeneluciferase construct. Growth medium for hepg2 dmem 10% fbs penstrep 1x procedure a. Hep2 is a cell line derived from hela they were are contamination in an originally hepatic cell line see atcc for details and so are a cervix carcinoma cell line, most useful, because they have a rather big nucleus. Growdexalginate hydrogel used for 3d coculture of hepg2.
This study aims to identify the active compounds of isf1 for the induction of follistatin expression. Complete growth medium the base medium for this cell line is atcc formulated eagles minimum essential medium, catalog no. No significant differences in hepg2 cell proliferation, adhesion, and albumin secretion were. Stable expression and integrated hepatitis b virus genome in. Culture of hepg2 liver cells on three dimensional polystyrene. Hepg2 cells have epithelial morphology and secrets a lot of plasma proteins. Moreover, apoptosis was assessed by measuring caspase3 activity. Hepg2 liver hepatocellular carcinoma cell culture and. Atcc is collaborating with the hcmi to offer scientists a wide variety of nextgeneration 2d and 3d patientderived in vitro cancer models, including organoids. Atcc c3a hepg2c3a, derivative of hep g2 atcc hb8065. Refer to the certificate of analysis for batchspecific test results. Nevirapine induced mitochondrial dysfunction in hepg2.
In this chapter, freezing, thawing, and subculturing procedures for hepg2 cells are described. Hepg2 is a human hepatoma that is most commonly used in drug metabolism and hepatotoxicity studies. Cell viability of hepg2 cells grown under hypoxic conditions declined to 81% from 6 to 72 h compared to normoxic conditions table 1. Table 1 proteins with altered expression in hepg2 cells after nevirapine treatment. Materials and methods cell culture hepg2 cells atcc hb 8065 were obtained from the american type culture collection manassas, va. Hepg2 cells are hepatocellular carcinoma hcc cells or hepatoblastoma cells. In vitro cell systems together with omics methods represent promising alternatives to conventional animal models for toxicity testing. Druginduced liver injury dili is a leading cause of discontinuation of new drug approval or withdrawal of marketed medicine based on safety due to organ vulnerability. However, whether sorafenib could be used to suppress the emt of hcc after insufficient rfa and further prevent the progression of residual hcc remains poorly unknown. Interferoninduced protein 44like ifi44l gene is a type i interferon. Gpc3, hspg and sulfatase2 sulf2 levels were measured by elisa. Highthroughput transcriptomic sequencing is an advanced approach for assessing the functional state of a tissue or cell type. Download fulltext pdf hep g2 is a hepatoblastomaderived cell line article pdf available in human pathology 4010.
Differentiated heparg cells cryopreserved description and. However, the influence of experimental factors, such as the sample preparation method and interlaboratory variation, on the transcriptomic profile has not been evaluated. Mcf7 breast adenocarcinoma national cancer institute. Receipt of frozen cells and starting cell culture 1 immediately place frozen cells in liquid nitrogen freezer storage until ready to culture. Transcriptomic and proteomic approaches have been widely applied in vitro but relatively few studies have used metabolomics. Huang a, dong j, li s, wang c, ding h, li h, su x, ge x, sun l, bai c, shen x, fang t, li j, shao n. Put an end to doityourself banking with each sixpack of readytouse quality control strains in glycerol stock. Atcc c3a hepg2 c3a, derivative of hep g2 atcc hb8065, liver tissue, frozen manufacturer. Hepg2 cells were treated either with oleic acid or tumor necrosis factor. Hepg2 is a widely used cell line in experiments of 30 years. Characterization of insulinlike growth factor ii receptors in human small cell lung cancer cell lines.
The human hepatocellular carcinoma cell line hepg2 atccnumber hb8065. Cells were fixed, embedded in paraffin wax, sectioned 10 m and. Osteopontindependent cd44v6 expression and cell adhesion. However, data on its physiological functions are limited and inconsistent. Given the there are many formulations of dmem and different versions of serum, we would like to provide the catalog numbers of the stuff that we are using. Human hepatoma hepg2 cells are used as an in vitro model of the human liver. Materials and methods cell culture hepg2 cells atcc. To make the complete growth medium, add the following components to the base medium. Growdexalginate hydrogel used for 3d coculture of hepg2 and. Sorafenib suppresses the epithelialmesenchymal transition of. The cells express 3hydroxy3methylglutarylcoa reductase and hepatic triglyceride lipase activities. High level expression of apoptosis inhibitor in hepatoma. Cell line was derived from the hepatocellular carcinoma cell line, hepg2 atcc hb8065. Warm emem media with 10% dialyzed fetal bovine serum prepared according to manufacturers recommendations in warm water bath for.
Cytotoxic effects of antiglypican3 against hepg2 cell line. Brightfield micrographs showing the structure of hepg2 cells cultured for 7 days on 22 mm diameter alvetex discs presented in the 6well plate format. The cells were grown as monolayer cultures in dulbeccos modified eagles medium dmem with the addition of 10% fetal. Epithelialmesenchymal transition emt played an important role in the progression of hepatocellular carcinoma hcc after insufficient radiofrequency ablation rfa. Table of contents this guide contains general technical information for bacterial growth, propagation, preservation, and application. Exosomal transfer of vasorin expressed in hepatocellular.
Furthermore, sorafenib inhibited the expression of pakt and perk12 in hcc cells in a dosedependent manner figs. Download highres image 479kb download fullsize imagefig. I was wondering as hepg2 is the cancer cell line fcs would be the better option to choose over fbs in terms of. The heparg cell line, a superior in vitro model to l02. Journal of nanomaterials hindawi publishing corporation. Hepg2 stable cell line generation and rnai applications sirna transfection. The human hepatocellular carcinoma cell line hepg2 atccnumber hb.
Nevirapine induced mitochondrial dysfunction in hepg2 cells. Surgical sutures coated with growdexalginate hepg2 and sewn three times through a pig liver segment indicating intact suture coating and live hepg2 cells within. Hepg2 cells are nontumorigenic cells with high proliferation rates and an epitheliallike morphology that perform many differentiated hepatic functions. Media is mem with 10% fbs and other stuff recommended, including ps. Surgical sutures coated with growdexalginatehepg2 and sewn three times through a pig liver segment indicating intact suture coating and live hepg2 cells within. Hepg2 cells were reversetransfected with scrna 200 nm or mir195 200 nm. This is a notimelimit, fully functioning version of the sim, but without sound communications with. Both cell lines have been maintained in our laboratory for more than 8 years.
The cells from passages 4 to 10 were used for the subsequent experiments. Cancer stem cells cscs have been considered to be one of the main mechanisms for chemoresistance, metastasis, and recurrent disease. Applic on n hepg2 labeling and cell growth protocol for. The hepg2 cell line is commonly used in drug metabolism and hepatoxicity studies. Apolipoprotein o apoo is a new member of the apolipoprotein family. Maintain the cells in the incubator to enable the 3d culture to become. Exosomal transfer of vasorin expressed in hepatocellular carcinoma cells promotes migration of human umbilical vein endothelial cells. Cellular morphology, proliferation, adhesion ability, and in vivo tumor formation were studied in the human hepatocellular carcinoma cell line hepg2 in the 3d rada16i scaffold. Dec 15, 2014 human hepatoma hepg2 cells are used as an in vitro model of the human liver. P urchase hepg2 cells b8065 as afroz en cll lt re m atcc m nassas, v.
You can use the pdf file attached for more details. The mrna level of pepck was analyzed at 48 h after reversetransfection by both rtpcr a and qrtpcr b. Liver cancer cell lines treated with doxorubicin under. Dorschner ordering information hepg2 can be ordered from atcc as a frozen ampoule. Active compounds in isf1 responsible for induction of follistatin were identified. C3a hepg2c3a, derivative of hep g2 atcc hb8065 atcc.
Hepg2 liver hepatocellular cells description, culturing medium conditions, transfection efficiency and protocols, and optimized transfection kits. Hepg2 cells red within the threads grew individually or in very small clusters. Sorafenib suppresses the epithelialmesenchymal transition. Costus speciosus is a medicinal plant commonly known as wild ginger distributed in south and southeast asian countries.
Pdf determining population doubling time and the appropriate. Hepatoblastoma hb or hepatocellular carcinoma is an embryonal malignancy of hepatocellular origin and the most common primary liver tumour of childhood, usually presenting in the first year of life. Hepg2 cells are useful as a model system in vitro for human hepatic cancer cells. Mar 30, 2016 druginduced liver injury dili is a leading cause of discontinuation of new drug approval or withdrawal of marketed medicine based on safety due to organ vulnerability. Using a microarray expression analysis, this study explored the function of apoo in liver cells. Culture and functional characterization of human hepatoma. This is a notimelimit, fully functioning version of the sim, but without sound communications with text messages only. These cells are epithelial in morphology, have a modal chromosome number of 55, and are not tumorigenic in nude mice. Data on the decreased expression of foxo1 by mir1271 in. Stable expression and integrated hepatitis b virus genome.
A human liver tumor cell line used to study a variety of liverspecific metabolic functions. This is the complete description of the cell line that you selected. Leaves of this plant are used for ayurvedic treatment regimes in malignancies and mental illness. The hepg2 liver cancer cells and chang normal liver cells were purchased from american type culture collection atcc, usa and cultured in dmem supplemented with 10% fbs, 100 gml streptomycin and 100 unitsml penicillin and maintained at 37 c with 5% co2 in humidified atmosphere. Differentiated heparg cells cryopreserved description and user guide thawing, culture and use 511 headquarters. Hepg2 is an immortalized cell line consisting of human liver carcinoma cells, derived from the liver tissue of a 15yearold caucasian male who had a welldifferentiated hepatocellular carcinoma, which is the fifth mostcommon cancer worldwide. Santamarine inhibits nfb and stat3 activation and induces. Hepatocellular carcinoma hcc is the second leading cause of cancerrelated death worldwide. Additional information on bacterial culturing can be found in bergeys manual of systematic bacteriology, 2nd edition, published by springer, new york1. Phase contrast micrographs of hepg2 cells grown in. Hepg2 cell proliferation was measured by mtt and lactate dehydrogenase ldh assays. Prepare a singlecell suspension of cells in media using trypsin to remove cells from 2d plasticware.
Such systems should ideally provide conditions that resemble those of natural host cells, e. Human hepg2 hepatic carcinoma cells were purchased from the american type culture collection atcc, manassas, vc, usa. The disease recurrent rate is relatively high resulted in poor 5year survival in advanced hcc. Hepg2 cell line is derived from a human hepatocellular carcinoma line we et. Rhizome extract from the plant is used to treat malignancies, pneumonia, urinary disorders, jaundice, rheumatism, and diabetes. Hep g2 is an immortal cell line which was derived from the liver tissue of a 15yearold african american adolescent boy with a welldifferentiated hepatocellular carcinoma. When cultured properly, they differentiate, forming apical and basolateral cell surfaces, resembling in vivo liver structures. Atcc c3a hepg2c3a, derivative of hep g2 atcc hb8065, liver tissue, frozen. This differentiation makes them very helpful in the study of intracellular transport of molecules between the different liver cell types, dysfunction of which can. The aim of this study was determining the viability and the growth rate of. An untargeted multitechnique metabolomics approach to. We report the use of the rada16i scaffold to mimic the ecm microenvironment and support tumor cell adherence and survival.
Microarray analysis provides new insights into the. Here, we investigated the role of foxa1 in the development of hcc by applying gene function gain and loss analysis to hepg2 and hep3b cell lines, and comparing outcomes with those of. Mcf7 breast adenocarcinoma ncipbcfhtb22 atcc htb22 february 27, 2012. We constructed a new plasmid, psehflaghbv, and transfected it into hepg2 cells, and. Cell culture growth conditions,nucleic acid purification,rnai.
Therefore, the goal of the present study was to develop an untargeted methodology for performing reproducible metabolomics. Osteopontindependent cd44v6 expression and cell adhesion in. No significant differences in hepg2 cell proliferation, adhesion, and albumin. Investigations of virushost interactions rely on suitable in vitro cell culture systems that efficiently support virus infection. Data on the decreased expression of foxo1 by mir1271 in hepg2 hepatocytes. In these cells, the hbv genome is integrated in several sites of hepg2 cellular dna.
I seeded them at 1x106100mm plate, and subculture in about 5 days without changing media during culture. Moreless the cells demonstrate decreased expression of. The aim of this research is to investigate the potential abilities of four different in vitro cell models l02, hepg2, heparg, and hiheps cell lines in assessing marketed drugs labeled with apparently different types of. To test the efficacy of kras inhibition in sorafenib resistance, we further established sorafenibresistant sr hep3b and hepg2 cells by using increasing doses. The chinese medicine formulation isf1 also known as buyanghuanwutang for poststroke rehabilitation could increase the expression of growthregulating protein follistatin by approximately 4fold. Alternatively, you can download the file locally and open with any standalone pdf reader. Atcc minis are authentic atcc cultures in a glassfree mini cryovial allowing you to get your assays moving faster just open, plate, and go.
Atcc crl10741 c3a is clonal derivative of hep g2 that was selected for strong contact inhibition of growth, high albumin production, high production of alpha fetoprotein afp and ability to grow in glucose deficient medium. Aixue huang, jie dong, shaohua li, chaonan wang, hongmei ding, hui li, xueting su, xingfeng ge, leqiao sun, chenjun bai, xuelian shen, tao fang, jie li. Expand populations of hepg2 cells in emembased media supplemented as described above as monolayer cultures in conventional 2d plasticware according to standard procedures. These multiple copies may have some influence on the cellular processes. Foxa1 inhibits hepatocellular carcinoma progression by. Hepg2h and smmc7721h cells showed significantly increased expression of pakt and perk12 compared with hepg2 and smmc7721 cells respectively figs. Apoptotic and inhibitory effects on cell proliferation of.
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