Female C57Bl/6 mice were purchased from Taconic Europe A/S (Denmark), and maintained under standardized conditions. Female Balb/c mice were purchased from JANVIER LABS (France). The mice were routinely used at the age of 8 to 12 weeks. All studies were approved by the Bioethics Committee in Lund, Sweden (M60-10 and M450-12), the Bioethics Committee of IPSEN (C2EA-13-048-S1) and by the French ministry (00617.01). All the experiments were conducted in accordance with the NIH Guide for the Care and Use of Laboratory Animals. Mice were treated with tasquinimod (30 mg/kg) ad lib. via drinking water or in the case of time studies by p.o. administration. The 4 T1 tumor cells were purchased from ATCC (France). The MC38 tumor cell line was obtained from S. A. Rosenberg (Nat. Cancer Inst., Bethesda, Maryland) and was transfected with the C215 antigen as previously described [52 ]. The cells were cultured in RPMI-1640 supplemented with ultra-glutamine (BioWhittaker/Lonza, Wokingham, UK); 10 % fetal bovine serum (Fisher Scientific, Pittsburgh, PA), 1 mM sodium pyruvate, 10 mM HEPES, 0.2 mg/ml gentamicine sulfate and 50 μM β-mercaptoethanol (R10 medium).
4t1 tumor cells
Manufactured by American Type Culture Collection
Sourced in United States
4T1 tumor cells are a commonly used murine mammary carcinoma cell line derived from a spontaneously arising tumor in a BALB/c mouse. The 4T1 cell line is highly tumorigenic and metastatic, making it a valuable model for studying breast cancer progression and metastasis.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Lonza (1 mentions)
Ultraglutamine, by Lonza (1 mentions)
Female balb c mice, by Janvier Labs (1 mentions)
Dulbecco modified eagle medium (dmem), by Fujifilm (1 mentions)
Penicillin streptomycin, by American Type Culture Collection (1 mentions)
Penicillin streptomycin, by Nacalai Tesque (1 mentions)
C57bl 6, by Charles River Laboratories (1 mentions)
Balb c mice, by Charles River Laboratories (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Dmem f12 media, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Zhejiang Tianhang Biotechnology (1 mentions)
Female balb c mice, by Charles River Laboratories (1 mentions)
6 protocols using 4t1 tumor cells
1
In Vivo Tumor Studies in Mice
2
Cell Culture Protocols for Cancer Cell Lines
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HeLa cells were purchased from the Human Science Research Resources Bank (Sennan‐shi, Japan) and maintained in DMEM (Wako) containing 10% FBS (Sigma‐Aldrich) and penicillin–streptomycin (Nacalai Tesque, Kyoto, Japan) at 37°C in a humidified atmosphere containing 5% CO2.
Lewis lung carcinoma (3LL) cells were maintained in RPMI‐1640 medium containing 10% FBS and penicillin–streptomycin at 37°C. 4T1 tumor cells were purchased from ATCC (Manassas, VA, USA) and cultured with RPMI‐1640 medium containing 10% FBS and penicillin–streptomycin at 37°C.
Lewis lung carcinoma (3LL) cells were maintained in RPMI‐1640 medium containing 10% FBS and penicillin–streptomycin at 37°C. 4T1 tumor cells were purchased from ATCC (Manassas, VA, USA) and cultured with RPMI‐1640 medium containing 10% FBS and penicillin–streptomycin at 37°C.
3
Anti-tumor Efficacy of QHL Compounds
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Example 8
Test purpose: to investigate the anti-tumor efficacy of QHL-086, QHL-087, QHL-092, QHL-095 in mice model for tumor treatment.
Test drug: QHL-086, QHL-087, QHL-092, QHL-095 and EMC-AANL-DOX injections and mitomycin injection, diluted to corresponding concentrations by physiological saline when testing.
Method and Results:
1. Animal: C57 mice of 6-8 weeks old, all female.
2. Production of tumor model
1) 4T1 tumor cells were purchased from American type culture collection (ATCC) and identified according the specification provided by ATCC. Cells were cultivated in DMEM culture solution containing 10% fetal bovine serum at 37° C. and 5% CO2. The cells were passaged for every three days and cells within the 15th passage were used.
2) Production of tumor. 5×106 4 T1 cells were subcutaneously injected to the back of the nude mice. Mice were randomly grouped after the tumor reached at least 100 mm3. Then treatment began and the day on which the treatment began was day 1.
3) Course of Treatment
According to the clinical application of QHL-086, QHL-087, QHL-092, QHL-095 and EMC-AANL-DOX were intravenously injected (IV) in a same dose of 36 umol/kg. The control group was administered by physiological saline. Drugs were administered once weekly for 3 weeks.
4) Results and discussions: As shown in
4
Syngeneic Murine Tumor Models
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MC-38 and A20 tumor cells were kindly provided by Yangxin Fu at Institute of Biophysics, Chinese Academy of Sciences (Beijing, China). The 4T1 tumor cells were purchased from the American Type Culture Collection (Manassas, Virginia, USA). Cells were cultured in Roswell Park Memorial Institute-1640 (RPMI-1640) or Dulbecco's Modified Eagle Medium (DMEM) medium with 10% fetal bovine serum in a humidified atmosphere with 5% CO2 at 37℃. Female C57/BL6 and BALB/c mice of 5 to 6 weeks age were purchased from Vital River Laboratory Animal Technology Co, Ltd (Beijing, China). To prepare syngenetic tumor models, A20 or 4T1 tumor cells (1×106 cells) were inoculated subcutaneously into the right flank of BALB/c mice, and MC-38 tumor cells (1×106 cells) were inoculated subcutaneously into the right flank of C57/BL6 mice.
5
Culturing 4T1 Breast Cancer Cells
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4T1 tumor cells, a mammary gland tumor cell line with autonomous metastatic potency, were purchased from American type culture collection (ATCC; Manassas, VA, USA) and cultured in Dulbecco's modified Eagle medium: nutrient mixture F-12 (DMEM/F12) media (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (Zhejiang Tianhang Biotechnology, Huzhou, Zhejiang, China), 100 IU/mL penicillin, and 100 μg/mL streptomycin (Gibco, Grand Island, NY, USA) in a humidified atmosphere of 5% CO2 at 37°C.
6
Orthotopic 4T1 Breast Cancer Metastasis in Mice
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Seven- to eight-week-old BALB/c female mice (obtained from Charles River Laboratories) were orthotopically inoculated into the right mammary fat pad with 1 × 104 viable 4T1 tumor cells (obtained from American Type Culture Collection) cultured according to previously described protocol [9 ]. Healthy BALB/c mice were used as a control group. Analysis was conducted 6 weeks after cancer cell transplantation. Animals (n = 5 for both control mice and mice with the cancer metastasis) were anesthetized by an intraperitoneal injection (i.p.) of a mixture consisting of ketamine and xylazine (100 mg ketamine/10 mg xylazine/kg body weight). The chest was opened and the thoracic aorta was quickly removed and transferred into Krebs–Henseleit buffer.
All experimental procedures involving animals were conducted according to the Guidelines for Animal Care and Treatment of the European Communities and the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised 1996). All procedures were approved by the Local Ethical Committee on Animal Experiments.
All experimental procedures involving animals were conducted according to the Guidelines for Animal Care and Treatment of the European Communities and the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised 1996). All procedures were approved by the Local Ethical Committee on Animal Experiments.
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