HUVECs were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured in endothelial growth medium (EGM-2), supplemented with the EGM-2-MV bullet kit (Lonza, Walkersville, MD, USA). The human lung adenocarcinoma A549 cell line and normal human bronchial epithelial cells were purchased from the Cell Resource Center of Life Sciences (Shanghai, China) and cultured in Dulbecco's Modified Eagle's medium (DMEM) containing 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Waltham, MA, USA). The Lewis lung carcinoma (LLC) cells and NCI-H1975 cells were purchased from ATCC and cultured in DMEM with 10% FBS. All media used contained 100 IU/ml penicillin and 100 μg/ml streptomycin. The cells were incubated in a humidified atmosphere of 5% CO2 at 37°C.
Lewis lung carcinoma cells
Manufactured by American Type Culture Collection
The Lewis Lung Carcinoma cells are a cell line derived from a spontaneous lung carcinoma in a C57BL/6 mouse. The cells are commonly used in cancer research to study tumor growth and metastasis.
Automatically generated - may contain errors
Lab products found in correlation
Crl 1642, by American Type Culture Collection (2 mentions)
Cd11c microbeads, by Miltenyi Biotec (2 mentions)
Nci h1975 cells, by American Type Culture Collection (1 mentions)
Huvecs, by American Type Culture Collection (1 mentions)
Egm 2mv bulletkit, by Lonza (1 mentions)
Raw 264.7 cells, by American Type Culture Collection (1 mentions)
Hela cell, by American Type Culture Collection (1 mentions)
Amicon ultra filtration columns, by Merck Group (1 mentions)
K562 cell, by American Type Culture Collection (1 mentions)
Ht29 human colorectal cancer cells, by American Type Culture Collection (1 mentions)
Primary human hscs, by ScienCell (1 mentions)
Mycoalert detection kit, by Lonza (1 mentions)
B16 melanoma cells, by American Type Culture Collection (1 mentions)
8 protocols using lewis lung carcinoma cells
1
Cell Culture Protocols for Cancer Research
2
Cell Culture Protocol for Diverse Cell Lines
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Primary antibodies, secondary antibodies, and affinity purified Fc-specific Fab fragments, as described in Table S1, were obtained from commercial sources. Any preservative, such as sodium azide or glycerol, was removed from the commercially obtained antibodies using 50 kDa molecular weight cut-off Amicon ultra filtration columns (Millipore, # UFC505096) and buffer exchanged into phosphate-buffered saline (PBS) buffer pH 7.2. Synthetic oligonucleotides (oligos) were obtained from IDT (Coralville, Iowa). Oligonucleotide sequences are presented in Table S2.
Cell culture RAW264.7 cells (mouse), Lewis Lung Carcinoma (LLC) cells (mouse), K562 cells (human) and HeLa cells (human) were obtained from ATCC (Manassas, VA) and were cultured as per manufacturer's recommendations. K562 CD11b/CD18 cells and mouse podocytes have been previously described (28, 29) (link). All cell lines were maintained according to the published procedures at 37°C with 5% CO2.
Cell culture RAW264.7 cells (mouse), Lewis Lung Carcinoma (LLC) cells (mouse), K562 cells (human) and HeLa cells (human) were obtained from ATCC (Manassas, VA) and were cultured as per manufacturer's recommendations. K562 CD11b/CD18 cells and mouse podocytes have been previously described (28, 29) (link). All cell lines were maintained according to the published procedures at 37°C with 5% CO2.
3
Murine and Human Hematopoietic Stem Cell Isolation
4
Generation and Characterization of Murine Tumor Cell Lines
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BrafV600EPten−/−(BPD6, male) cell lines were generated and cultured in DMEM with 10% FBS at 37°C as previously described (Holtzhausen et al., 2015 (link)). KrasG12Dp53flox/flox cell lines were generated through enzymatic and mechanical digestion of autochthonous mouse lungs during week 16 post-virus administration by visual tumor dissection and serial culturing in DMEM with 10% FBS to remove fibroblasts. Cell lines were confirmed by rt-PCR and western blot. Lewis Lung Carcinoma cells were purchased from the ATCC (ATCC, CRL-1642) and cultured according to the supplier’s specifications. Bone marrow-derived dendritic cells (BMDCs) were harvested and differentiated using IL-4 (BioAbChem, 42-IL4 C) and GM-CSF (BioAbChem, 42-GMCSF) as previously described and purified using CD11c microbeads (Miltenyi Biotec, 130-108-338) according to the manufacturer’s protocol (Inaba et al., 1992 (link)). The HEK293-LEF1/TCF-luciferase cell line was cultured in DMEM with 10% FBS as previously described in the presence or absence of OMP-18R5 or OMP-54F28 (Ring et al., 2011 (link)). The murine dendritic cell line, DC2.4 (a generous gift from Dr. Kenneth Rock, University of Massachusetts Medical School) was cultured and generated as previously described (Shen et al., 1997 (link)).
5
Generation and Characterization of Murine Tumor Cell Lines
6
Genetically Modified Mouse Model for Lung Cancer
7
Murine Breast and Lung Cancer Models
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The polyoma middle T (PyMT) murine breast carcinoma cell line was derived from the primary tumor-bearing transgenic mice expressing PyMT under the control of the murine mammary tumor virus promotor [15 (link)]. These cells were kindly provided by Prof. Tsonwin Hai (Ohio State University, Columbus, OH, USA). The Lewis lung carcinoma (LLC) cells were purchased from the American Type Culture Collection (ATCC). Cells were tested and found to be free of mycoplasma contamination. Cells were cultured in DMEM supplemented with 10% fetal bovine serum (FBS), 1% streptomycin and penicillin at 37 °C, in a humidified atmosphere containing 5% CO2.
PyMT and LLC cancer cells (106) were implanted into the mammary fat pad of female mice or subcutaneously into the flanks and orthotopically injected into male mice, respectively. Tumor volume was monitored over time with a caliper and calculated using the formula: width2 × 0.5 × length. According to the Institutional Animal Care and Use Committee, the humane endpoint is defined when the tumor size reaches 1500 mm3. Tumors of PyMT and LLC cells were maintained for 25 and 20 days, respectively, following the cancer cells injection. Both cancer cells models reached similar tumor weight at the endpoint.
PyMT and LLC cancer cells (106) were implanted into the mammary fat pad of female mice or subcutaneously into the flanks and orthotopically injected into male mice, respectively. Tumor volume was monitored over time with a caliper and calculated using the formula: width2 × 0.5 × length. According to the Institutional Animal Care and Use Committee, the humane endpoint is defined when the tumor size reaches 1500 mm3. Tumors of PyMT and LLC cells were maintained for 25 and 20 days, respectively, following the cancer cells injection. Both cancer cells models reached similar tumor weight at the endpoint.
8
Lewis Lung Carcinoma Cell Culture
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The Lewis Lung Carcinoma (LLC) cells were purchased from the American Type Culture Collection (Manassas, VA) and were cultured at 37°C in 5% CO2 −95% air using Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum.
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