Human NIH:OVCAR-3 cell was purchased from the Bioresource Collection and Research Center (Hsinchu, Taiwan). TE1 and TE9 cells, originally purchased from the Japanese Collection of Research Bioresources, were kindly provided by Dr. Yann-Jang Chen (National Yang-Ming University, Taiwan). Ishikawa cell, originally purchased from the European Collection of Cell Culture, was kindly provided by Dr. Sin-Tak Chu (Academia Sinica, Taiwan). Other human cell lines, including HEK-293T, THP-1, KATO III, T-47D, RL95-2, HEC-1-A, HeLa and SKOV-3, were originally from ATCC.
Rl95 2
Manufactured by American Type Culture Collection
Sourced in United States
The RL95-2 is a cell line derived from human endometrial carcinoma. It is a widely used in vitro model for the study of endometrial cancer. The RL95-2 cell line maintains characteristics of well-differentiated endometrial adenocarcinoma and is commonly utilized for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Hec 1a, by American Type Culture Collection (42 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (30 mentions)
Hec 1b, by American Type Culture Collection (25 mentions)
An3ca, by American Type Culture Collection (24 mentions)
Ishikawa, by American Type Culture Collection (16 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (11 mentions)
Rl95 2, by Thermo Fisher Scientific (10 mentions)
Dmem f12, by Thermo Fisher Scientific (9 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (9 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (8 mentions)
Hec 1a, by Thermo Fisher Scientific (7 mentions)
An3ca, by Thermo Fisher Scientific (5 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (5 mentions)
Ishikawa, by Thermo Fisher Scientific (5 mentions)
Hec 1b, by Thermo Fisher Scientific (4 mentions)
Rpmi 1640, by Thermo Fisher Scientific (4 mentions)
Ishikawa, by Merck Group (3 mentions)
Insulin, by Merck Group (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Mcf 7, by American Type Culture Collection (3 mentions)
89 protocols using rl95 2
1
Cell Line Procurement for Cancer Research
2
Endometrial Cancer Cell Line Engineering
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Ishikawa cell, originally purchased from the European Collection of Cell Culture, was kindly provided by Dr. Sin-Tak Chu. RL95-2 and HEC1A cells were originally obtained from ATCC. For gene manipulation, the cDNA encoding eGFP or human NMUR2 was amplified and subcloned into the pLAS2w.Ppuro lentivector. The shGFP-expressing lentivector, shNMU-expressing lentivector, shCD44-expressing lentivector and other plasmids used for the lentivirus production and establishment of stable cell lines were purchased from National RNAi Core Facility Platform, Taiwan. Infected endometrial cancer cells were selected and maintained in the puromycin-containing medium.
3
Establishing Human EC Cell Line Cultures
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Human EC cell lines (Ishikawa, HEC-1A, KLE, ECC-1, AN3-CA, and RL95-2) were purchased from ATCC and maintained in Dulbecco’s Modified Eagle Medium (DMEM) /F12 (Gibco, Carlsbad, CA, USA) and RPMI-1640 medium (Gibco) containing 10% fetal bovine serum (Gibco) and 100 U/mL of penicillin/streptomycin in a humidified incubator of 5% CO2 in compressed air at 37 °C. We purchased all reagents from Sigma-Aldrich unless stated otherwise.
4
Establishing Endometrial Cancer Cell Lines
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Commercial endometrial cancer cell lines, i.e. MFE-296 cells were purchased from DSMZ GmbH (Berlin, Germany) and cultured in MEM with 10% fetal bovine serum (FBS) and 1X penicillin/streptomycin (Pen/Strep). SNG-M cells were purchased from the JCRB cell bank (Osaka, Japan) and cultured in Ham’s F12 median with 10% FBS and 1X Pen/Strep. HEC1A (McCoy’s 5a with 10% FBS and 1X Pen/Strep), RL95-2 (DMEM:F12, 0.005 mg/mL insulin, 10% FBS and 1X Pen-Strep), Ishikawa (MEM with 2 mM Glutamine, 5% Fetal Bovine Serum (FBS) and 1X Pen/Strep), AN3CA (EMEM with 10% FBS and 1X Pen/Strep ) and KLE (DMEM:F12 with 10% FBS and 1X Pen/Strep) cells were from ATCC (Manassas, VA). NCI-EC1, ACI-80, ACI-181, ACI-52, ACI-61 and ACI-68 were obtained from John Risinger at Michigan State University (East Lansing, MI) and maintained in DMEM:F12 with 10% FBS and 1X Pen/Strep (ATCC). Uterine leiomyoma tissues (ULM, discovery cohort), Age-matched, formalin-fixed, paraffin embedded uterine leiomyomas from self-described white and black women were obtained under an institutional-review board approved protocol from Inova Fairfax Hospital (Falls Church, VA). Uterine leiomyoma tissues (ULM, validation cohort), flash-frozen uterine leiomyomas were obtained under an institutional-review board approved protocol from Johns Hopkins Memorial Institute (Baltimore, MD).
5
Endometrial Carcinoma Cell Line Cultivation
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Endometrial carcinoma cell lines KLE (ATCC CRL1622TM), AN3CA (ATCC HTB111TM), RL-95-2 (ATCC CRL1671TM), and human fibroblast (ATCC CRL-2106) were purchased from ATCC (American Type Culture Collection, Manassas, VA, USA). Ishikawa was purchased from Sigma-Aldrich (St. Louis, MO, USA). AN3CA and fibroblast cell lines were maintained in MEM (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) and 2% penicillin/streptomycin (PAN-Biotech GmbH, Aidenbach, Germany). The Ishikawa cell line was maintained in MEM supplemented with 5% FBS and 2% penicillin/streptomycin. KLE cell line was maintained in DMEM (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS and 2% penicillin/streptomycin. RL-95-2 cell line was maintained in DMEM: F12 (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS, 2% penicillin/streptomycin, and 0.005 mg/mL insulin (Sigma-Aldrich, St. Louis, MO, USA). All cells were grown in a humidified chamber at 37 °C in 5% CO2 atmosphere.
6
Cultured Cell Lines Treatment
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Ishikawa 3-H-12 (IK), HEC-1-A, RL95-2 and KLE cells were purchased from the ATCC (Manassas, VA, US). The four cell lines were cultured in DMEM (Sigma-Aldrich, St Louis, MO, USA) supplemented with 10% FBS (Invitrogen, Barcelona, Spain), 20 mmol/L of L-Glutamine and penicillin/streptomycin. Sunitinib was kindly provided by Pfizer and LY294002 was purchased from Calbiochem (La Jolla, CA, USA). All treatments were carried out in complete media, performed in triplicate and repeated at least 3 times.
7
Comprehensive Panel of Human Cancer Cell Lines
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Human cancer cell lines NCI‐H1975 (non‐small‐cell lung cancer), 786‐O (renal cell adenocarcinoma), NCI‐N87 (gastric carcinoma), DU145 and PC‐3 (prostate carcinoma), RL95‐2 (endometrial carcinoma), BxPC‐3 (pancreatic adenocarcinoma), TF‐1α (erythroleukemia), MDA‐MB‐231 and MCF‐7 (breast adenocarcinoma), COLO205 (colorectal adenocarcinoma), CCRF‐CEM (acute lymphoblastic leukemia), CHO‐K1, and the murine myeloma cell line P3X63Ag8U.1 were purchased from ATCC (Manassas, VA, USA). NCI‐H322 (non‐small‐cell lung cancer) was purchased from European Collection of Cell Cultures (Salisbury, UK). HEC‐1 (endometrial carcinoma) was purchased from Japanese Collection of Research Bioresources (Osaka, Japan).
8
Characterization of Endometrial Cancer Cell Lines
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The following EC epithelial cell lines were used in this study. HEC-1A (Ref. HTB112, ATCC; Manassas, VA, USA) cell line was cultured in McCoys 5 A medium (Ref. 80014020, ThermoFisher; Waltham, MA, USA); Ishikawa (Ref. 99040201, Sigma-Aldrich; St. Louis, MO, USA) and RL95-2 (CRL-1671, ATCC; Manassas, VA, USA) were cultured in DMEM/F12 (Ref. 11320-033, ThermoFisher; Waltham, MA, USA). Both supplemented with penicillin/streptomycin (1%) and fetal bovine serum (10%). Incubated at 37 °C in a 5% CO2 humidified chamber. Cell lines were morphologically and genetically authenticated and tested for mycoplasma in accordance with AACR guides.
9
Metformin Treatment on Endometrial Cancer
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RL95‐2 was purchased from ATCC (Manassas, VA, CRL‐1671) and maintained in DMEM:F‐12 medium (ATCC 30‐2006) supplemented with 10% fetal bovine serum (ATCC 30‐2020), 100 U/mL penicillin/100 µg/mL streptomycin (ATCC ATCC 30‐2300), and 0.005 mg/mL insulin (Sigma‐Aldrich, I9278). ACI‐181 (RRID:CVCL_N828),18 a model of endometrioid endometrial cancer (EEC) was a gift from Dr John Risinger (Department of Obstetrics, Gynecology and Reproductive Biology, Michigan State University) and cultured in DMEM:F12 supplemented with 10% fetal bovine serum, 100 U/mL penicillin, and 100 µg/mL streptomycin. Both cell lines were maintained at 37°C and 5% CO2. Metformin was purchased from Tocris (2864). For the Metformin time course experiments, 100 000 and 200 000 of ACI‐181 and RL95‐2 were seeded in 6‐well plates and treated with 20 mmol/L of Metformin for 96 and 120 hours. Two independent experiments were performed with one biological sample for each treatment condition per cell line.
10
Endometrial Carcinoma Cell Line Protocol
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Human endometrial carcinoma cell lines ECC1, HEC-1A, HEC-1B, HEC-50, HEC-59, Ishikawa, MFE-296, MFE-280 and RL-952 (ATCC) were cultured in RPMI-1640 with 2 nM L-glutamine (Invitrogen, Carlsbad, CA, USA) and 10% fetal bovine serum. Human USC cell lines ARK1 and ARK2 were a kind gift from Dr. Alessandro D. Santin (Yale Cancer Center, New Haven, CT, USA), and USC cell lines ACI-126 and ACI-158 were a kind gift from Dr. John I. Risinger (Michigan State University, East Lansing, MI, USA). All four cell lines were cultured in DMEM with 1.0 g/L glucose, L-glutamine, sodium pyruvate (Corning Cellgro, Corning, NY, USA) and 10% fetal bovine serum. All cells were maintained at 37 °C in 5% CO2. Use of the cell lines in experiments were within the first 10 passages. All cells were tested negative for mycoplasma contamination using the MycoAlert mycoplasma detection kit (Lonza Group Ltd., Basel, Switzerland) at the time of thawing the frozen cell lines and were authenticated by short tandem repeat profiling in the Characterized Cell Line Core at the University of Texas MD Anderson Cancer Center.
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