The choriocarcinoma trophoblast cell lines, BeWo, Jar, and Jeg‐3, were from American Type Culture Collection (ATCC). The non‐tumor trophoblast HTR‐8/SVneo was a gift from Prof. Charles Graham (Queen's University at Kingston, Canada). Human umbilical vein endothelial cells (HUVECs; originally from ATCC) and aortic endothelial cells (HAoECs; originally from PromoCell GmbH) were provided by Dr. Andriana Margariti and Prof. David Grieve, respectively (Queen's University Belfast, UK). BeWo cells were maintained in DMEM/F12 Gibco (Thermo Fisher) supplemented with 2 mmol/L L‐glutamate. Jar and HTR‐8/SVneo were cultured in RPMI 1640 (Sigma‐Aldrich), and Jeg‐3 was cultured in EMEM (ATCC). Both HUVECs and HAoECs were cultured in EGM‐2 (Lonza) with all vendor‐provided supplements in flasks or plates coated with 0.1% gelatin (Sigma‐Aldrich). All growth media contained 10% fetal calf serum. There was no mycoplasma contamination with the cells.
Jeg 3
Manufactured by American Type Culture Collection
Sourced in United States, China
The JEG-3 is a human choriocarcinoma cell line derived from placental trophoblasts. It is a commonly used in vitro model for studying placental development and function.
Automatically generated - may contain errors
Lab products found in correlation
Htr 8 svneo, by American Type Culture Collection (19 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (16 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (12 mentions)
Dmem f12, by Thermo Fisher Scientific (8 mentions)
Rpmi 1640, by Thermo Fisher Scientific (7 mentions)
Hek293t, by American Type Culture Collection (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (6 mentions)
Mcf 7, by American Type Culture Collection (5 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (5 mentions)
Jeg 3, by Thermo Fisher Scientific (5 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions)
Minimum essential medium (mem), by Thermo Fisher Scientific (4 mentions)
L glutamine, by Thermo Fisher Scientific (3 mentions)
Huvecs, by American Type Culture Collection (3 mentions)
Rpmi 1640, by Merck Group (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Htr 8 svneo, by Thermo Fisher Scientific (3 mentions)
L glutamine, by Lonza (3 mentions)
Htb 144, by American Type Culture Collection (2 mentions)
84 protocols using jeg 3
1
Choriocarcinoma Trophoblast Cell Lines Protocol
2
Establishment of Human Endothelial Cell Lines
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Primary hBMECs (passage 1) isolated from human brain were purchased from ScienCell Research Laboratories (Santiago, CA, USA). HUVECs (passage 3) were purchased from the American Tissue Culture Collection (ATCC) (Manassas, VA, USA). Both hBMECs and HUVECs were cultured in endothelial cell basal medium supplemented with endothelial cell growth supplement, penicillin-streptomycin solution, and 5% fetal bovine serum (FBS) (Invitrogen) at 37°C in a 5% CO2 atmosphere. The human placenta choriocarcinoma cell line JEG-3, human lung adenocarcinoma cancer cell line A549, African green monkey kidney cell line Vero, and HEK293T cells were purchased from ATCC and cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with penicillin (100 U/ml), streptomycin (100 mg/ml), 2 mM glutamine, and 10% FBS and maintained at 37°C in 5% CO2.
3
Culturing Human Cell Lines for Transport Studies
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The human choriocarcinoma cell lines Jeg-3 and BeWo were purchased from ATCC and grown in Dulbecco’s Modified Eagle’s Medium (DMEM) and Ham’s F12K Kaighn’s Medium, respectively. HEK 293 cell lines transfected with P-gp, ABCG2, or MRP1 were grown in DMEM supplemented with 1 mg/mL geneticin35 (link). Drug-selected H460-MX20 cells that overexpress ABCG2 were grown in RPMI media containing 20 nM mitoxantrone. All media above was supplemented with 10% Fetal Bovine Serum (FBS), penicillin/streptomycin, and 2 mM glutamine. Primary human villous trophoblast (HVT) cells were purchased from Sciencell Research Laboratories and maintained in Trophoblast medium (Sciencell) with included supplements. All cells were incubated at 37 °C in 5% CO2. Cells were sub-cultured utilizing a 0.25% trypsin, 9 mM EDTA solution, and were used in experiments between passages 4–20.
4
Cultivation of Human Placental Cell Lines
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HTR-8/SVneo (ATCC CRL-3271, Manassas, VA, USA), an immortalized human first trimester placental cell line, was derived by transfection of first trimester human trophoblasts with a gene encoding simian virus 40 large T antigen. Choriocarcinoma-derived placenta trophoblast cell lines JEG-3 (ATCC HTB-36, Manassas, VA, USA) and BeWo cells (ATCC CCL-98, Manassas, VA, USA). The three cells were grown on RPMI1640 medium + Gluta MAX (Gibco; McKinley Place NE; MN; USA) combined with 10% fetal bovine serum (FBS, Gibco; McKinley Place NE; MN; USA). The cells were refined in a humid incubator at 37 °C under 5% carbon dioxide. Replacement of the cultural mediums occurred between 2 and 3 days. Cells were planted in each well of a 96-well, with saturation of cell viability and multiplication studies. ELISA testing, hormone measurement, and wound healing assays were all performed in 24-well plates. Cells were cultivated in 6-properly saturations to aid in Western blotting and for real-time polymerase chain reaction (RT-PCR).
5
Comprehensive Ovarian and Cervical Cancer Cell Lines
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The following cell lines were purchased from ATCC: CAOV-3 (ovarian adenocarcinoma; HTB-75), SK-OV-3 (ovarian adenocarcinoma; HTB-77), SW 626 (ovarian adenocarcinoma; HTB-78), ES-2 (ovarian clear cell carcinoma; CRL-1978), PA-1 (ovarian teratocarcinoma; CRL-1572), TOV-21G (ovarian clear cell carcinoma; CRL-11730), HeLa (cervical adenocarcinoma; CCL-2), Ca ski (cervical epidermoid carcinoma; CRL-1550), C-33 A (cervical carcinoma; HTB-31), JAR (placental choriocarcinoma; HTB-144), JEG-3 (placental choriocarcinoma; HTB-36), and HEK 293T (embryonic kidney fibroblast; CRL-3216).
6
Characterization of Diverse Cell Lines
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We have previously described iPSC lines derived from two MSCs, namely adipose stem cell (ASC; Invitrogen, Carlsbad, CA, USA) and human adipose-derived MSC (MSC-AT; PromoCell, Heidelberg, Germany), and from a human white pre-adipocyte (HWP) cell line [4 (link), 27 (link)]. In this work, human adipose-derived MSC, designated ASC Lonza, was purchased from Lonza, Lonza, Verviers, Belgium. MH#1 was an iPSC cell lined established from ASC Lonza in our lab (S. Sugii, unpublished data). WJ0706 is a human MSC cell line derived from Wharton’s Jelly (WJ) obtained from Cytopeutics Sdn. Bhd, Selangor, Malaysia (http://www.cytopeutics.com ). The MSC cell lines were isolated and characterized at Cytopeutics according to standard procedures and with ethical clearance [28 (link)]. Human placenta choriocarcinoma cell line JEG-3 (ATCC HTB-36), human normal placental cell line HS 799. PI (ATCC CRL-7530) and human normal colon cell line CRL-1790 (ATCC CRL-1790) were purchased from ATCC (Manassas, VA, USA). Cancer cell lines were kindly provided by Professor Y.M. Lim, Cancer Research Center, Universiti Tunku Abdul Rahman.
7
Cell Culture of Human Carcinoma Lines
8
Trophoblast Cell Line Stimulation Assay
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The choriocarcinoma cell lines BeWo (ECACC, Salisbury, UK) and JEG-3 (ATCC), which are useful models of human trophoblasts, were used for the study. The cells were cultured in DMEM (3.7 g/L NaHCO3, 4.5 g/L d -glucose, 1.028 g/L stable glutamine, and Na-Pyruvate; Biochrom, Berlin, Germany). 10% heat-inactivated FCS was added to the medium, and the solution was incubated at an atmospheric CO2 concentration of 5% and at 37°C.
BeWo and JEG-3 cells were separately grown on sterile 12 multiwell slides at a density of 500,000 cells/mL DMEM with 10% foetal cow serum. After 4 h, medium was changed to pure DMEM. The cells were stimulated with 0.01 nM or 0.1 nM triiodothyronine (T3) (Sigma T2877; Lot:106K1157V, Sigma-Aldrich), thyronamine (T0AM) (Fluka 80345, Lot: BCBL2185V, Buchs, Switzerland), 3-Iodothyronamine (T1AM) (Cayman Chemical) and RO5203548 (Glixx Laboratories, Hopkinton, MA, USA) for 2 h (PCR samples) and 48 h (Immunocytochemistry and Western blot samples), respectively. Control cells were incubated without stimulants. As a control solvent, the following buffers were used: The stimulant T3 has been diluted in 1 M NaOH in DMEM (Dulbecco’s Modified Eagle Medium) and T1AM and RO5203548 were diluted in DMSO (Dimethylsulfoxide).
BeWo and JEG-3 cells were separately grown on sterile 12 multiwell slides at a density of 500,000 cells/mL DMEM with 10% foetal cow serum. After 4 h, medium was changed to pure DMEM. The cells were stimulated with 0.01 nM or 0.1 nM triiodothyronine (T3) (Sigma T2877; Lot:106K1157V, Sigma-Aldrich), thyronamine (T0AM) (Fluka 80345, Lot: BCBL2185V, Buchs, Switzerland), 3-Iodothyronamine (T1AM) (Cayman Chemical) and RO5203548 (Glixx Laboratories, Hopkinton, MA, USA) for 2 h (PCR samples) and 48 h (Immunocytochemistry and Western blot samples), respectively. Control cells were incubated without stimulants. As a control solvent, the following buffers were used: The stimulant T3 has been diluted in 1 M NaOH in DMEM (Dulbecco’s Modified Eagle Medium) and T1AM and RO5203548 were diluted in DMSO (Dimethylsulfoxide).
9
Culturing Diverse Cell Lines and Bacterial Strains
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Human HeLa cells (ATCC CCL2), HEK293-FT (Invitrogen), JEG-3 (ATCC HBT-36), HepG2 (ATCC HB-8065) and LoVo cells (ATCC CCL229) were cultured at 37 °C under an atmosphere of 5% CO2, in media recommended by ATCC (Manassas, VA). The culture media (GIBCO) was supplemented with 10% Fetal Calf Serum (FCS) (Sigma). L. monocytogenes and Escherichia coli strains were grown in Brain–Heart Infusion (BHI) medium or Luria–Bertani (LB) medium (Difco, BD), respectively, with antibiotics in the presence of plasmids. Bacterial strains and plasmids are described in the Supplementary Information.
10
Genetic engineering of human cancer cells
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Human cancer cell lines NCCIT, NTERA-2 a1.D1, JAR, JEG-3, and SKBR3 were obtained from ATCC (CRL-2073, CRL-1973, HTB-144, HTB-36, and HTB-20). A549 and HeLa were obtained from RIKEN (RBRC-RCB0098, RBRC-RCB0007). PANC-1 was obtained from DS pharma biomedical (EC87092802-F0). All samples used were tested for mycoplasma contamination. We confirmed that our cell lines are negative to mycoplasma contamination. The piggyBac transposon vector carrying tetO-HA-Dmrt1/Venus-ires-mCherry-Ef1a-rtTA and pCAG-PBase was transfected into human cancer cell lines using Lipofectamine 2000 (Thermo Fisher Scientific). After antibiotic selection with 350 μg/mL G418, the surviving cells were expanded and used for analyses.
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