All animal protocols were in full compliance with the guidelines for animal care and were approved by the Animal Care Committee from the Animal Ethics committee of Huaqiao University. Six–to-eight weeks old male nude BALB/c mice (H-2b) were obtained from the Experimental Center of the Medical Scientific Academy of Fujian. Human umbilical vein endothelial cells (HUVECs, ATCC, CRL-1730) and the cervical cancer cell line HeLa (ATCC, CCL-2) were purchased from American Tissue Type Culture Collection. HUVECs were routinely cultured at 37°C in EGM-2 medium supplemented with 10% FBS. The HeLa cell line was cultured at 37°C in RPMI-1640 medium supplemented with 10% FBS.
Atcc ccl 2
Manufactured by American Type Culture Collection
Sourced in United States
ATCC CCL-2 is a continuous cell line derived from human epithelial cells. It is a commonly used model for various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (8 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions)
L glutamine, by Thermo Fisher Scientific (5 mentions)
Atcc htb 22, by American Type Culture Collection (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Mda mb 231, by American Type Culture Collection (2 mentions)
Mcf 7, by American Type Culture Collection (2 mentions)
Atcc crl 1573, by American Type Culture Collection (2 mentions)
Atcc htb 26, by American Type Culture Collection (2 mentions)
100 mm plastic tissue culture dishes, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Foetal bovine serum (fbs), by Merck Group (1 mentions)
Amphotericin b, by Merck Group (1 mentions)
Atcc ccl 1, by American Type Culture Collection (1 mentions)
Gentamycin, by Thermo Fisher Scientific (1 mentions)
Dulbecco, by Thermo Fisher Scientific (1 mentions)
Cas 7647 14 5, by Merck Group (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt rpmi1640, by Thermo Fisher Scientific (1 mentions)
Crl 3435, by American Type Culture Collection (1 mentions)
Silver nitrate agno3, by Merck Group (1 mentions)
59 protocols using atcc ccl 2
1
Nude Mouse Model for HUVEC and HeLa Cells
2
Cytotoxicity and ROS Evaluation in HeLa Cells
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Human cervical cancer cells (HeLa) (ATCC® CCL2™), obtained from ATCC, were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin (Gibco BRL, Grand Island, NY) and were maintained in a humidified incubator at 5% CO2 and 37°C. Cells were routinely grown in 100 mm plastic tissue culture dishes (Nunc, Roskilde, Denmark) and harvested with a solution of trypsin-EDTA, while in a logarithmic phase of growth. The CCK-8 and LDH assays were performed as described previously [25 (link), 26 (link)]. Cell membrane integrity of the human cervical cancer cells was evaluated by determining the activity of lactate dehydrogenase (LDH) leaking out of the cells, as per the manufacturer's instructions and also as described previously [25 (link)]. ROS were estimated according to a method described previously [27 (link)].
3
HeLa Cell Culture Protocol
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Cervical cancer, HeLa cells (adenocarcinoma, HPV18, ATCC® CCL2™) were directly procured from ATCC (American Type Culture Collection, Manassas, VA, USA). These cells were amply characterized and authenticated by the ATCC using PCR for viral genome sequencing and DNA profiling, cytogenetic analysis was also performed by the ATCC for a full description of the cytogenetic information of the HeLa cells. The cells were passaged at the Laser Research Centre, University of Johannesburg upon receipt following ATCC’s recommendations for thawing frozen vials. They were cultured in liquid medium, Minimum Essential Medium Eagle’s, MEME, (Sigma Aldrich: M2279). Complete media was prepared by supplementing MEME with 10% Foetal Bovine Serum, FBS, (Sigma Aldrich: F0804), 1% sodium pyruvate (), 2% L-glutamine (), 100 mg penicillin and streptomycin (Sigma Aldrich: P4333100ML), and 100 mg Amphotericin B (Sigma Aldrich: A2942-100ML). The cells were cultured as a monolayer in an incubator set at 37° C with 5% CO2 and 85% humidity.
4
Cell Culture Protocols for Cancer Research
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The A549 cell line were obtained from ECACC and maintained in DMEM or EMEM, respectively, supplemented with 2 mM of glutamine, 10% fetal bovine serum, and antibiotics (1% of 100 U/L penicillin, 100 mg/mL streptomycin). L929 (NCTC clone 929, ATCC® CCL-1™) and HeLa (ATCC® CCL-2™) cell lines were obtained from ATCC and maintained in EMEM supplemented with antibiotics (1% of 100 U/L penicillin, 100 mg/mL streptomycin) and with 5% and 10% FBS respectively. Cell line was routinely grown in tissue culture flasks (75 cm2) and kept in a humidified atmosphere of 5% CO2 at 37 °C. All examined cell lines was tested against mycoplasma contamination with microbiological assays. The research was carried out using the normal cell line L929—mouse fibroblast and cancer cells: HeLa—human cervical cancer, A549—human lung cancer. All of the stock solutions of the tested compounds were dissolved in DMSO.
5
Cell Line Culturing for Cancer Research
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Human breast adenocarcinoma (MDA-MB-231, ATCC® CRM-HTB-26™) cell line and HeLa human cervix cancer cells (ATCC® CCL-2™) were purchased from ATCC (Manassas, VA, USA). MDA-MB-231-Luc-RFP stable cell line was obtained from AMSBIO (SC041, Abingdon, UK). Cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (Invitrogen, Cergy Pontoise, France) and gentamycin (0.05 mg mL−1) (Invitrogen, Cergy Pontoise, France) and incubated at 37 °C in a humidified atmosphere with 5% CO2.
6
Transcription Enhancement via dCas9-VPR
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For enhancing sufficient levels of gene transcription, we selected dCas9-VPR, which fused with the three different types of cis-acting transcription activation domain (Supplementary Fig. 18a )25 (link). After confirming that the dCas9-VPR can activate the two different target genes, IL1RN and SLC28A2, with separate three sgRNAs, respectively, we generated a stable cell line expressing dCas9-VPR selected by G418 (500 μg/mL, Invitrogen) to the transfected Hela cells (ATCC® CCL-2™). The 3800 sgRNA libraries were stably overexpressed by infection of the lentiviral particles into the dCas9-VPR Hela to generate SLC-CRISPRa pools (Supplementary Fig. 18c ). The sgRNA expressing cells were selectively expanded by incubating the cells with puromycin (2 μg/mL). To maintain the diversity of the pools, >1.52 × 106 cells were plated for subsequent culture and used for the initial sorting with a fluorescent probe (Supplementary Fig. 18c ).
7
Isolation and Culture of Murine Macrophages
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Primary bone-marrow-derived macrophages (BMDMs) and peritoneal macrophages were isolated from male and female wild-type C57BL6/J mice. Bone marrow was harvested from both femurs, red blood cells were lysed and resulting marrow cells were cultured in 70% DMEM (10% v/v FBS, 100 U/mL penicillin, 100 μg/mL streptomycin) supplemented with 30% L929 mouse fibroblast-conditioned media for 6–7 days. BMDMs were seeded out the day before at a density of 1 × 106 mL−1 in DMEM (10% v/v FBS, 100 U mL−1 penicillin, 100 μg mL−1 streptomycin). Peritoneal macrophages were isolated by peritoneal lavage and seeded out overnight at a density of 1 × 106 mL−1 in DMEM (10% v/v FBS, 100 U mL−1 penicillin, 100 μg mL−1 streptomycin). HeLa cells were seeded out at 0.1 × 106 mL−1 in DMEM (10% v/v FBS, 100 U mL−1 penicillin, 100 μg mL−1 streptomycin). HeLa cells were obtained from ATCC (HeLa (ATCC CCL-2)) and are periodically tested for mycoplasma.
8
Doxorubicin Treatment of HeLa Xenografts
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The nude mice were inoculated subcutaneously with HeLa cells (ATCC® CCL-2™) to generate xenograft tumors. Post-inoculation, the mice were intraperitoneally treated with either doxorubicin (CAS 25316-40-9, Sigma-Aldrich, St. Louis, MO, United States) at varying doses between 2 and 3 mg/kg, or normal saline (CAS 7647-14-5, Sigma-Aldrich, St. Louis, MO, United States) at doses between 10 and 12 mg/kg, twice a week. The treatments started from the seventh day post-inoculation, and the experiment lasted for 28 days.
9
Antifungal Activity Screening of Fungi
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Twelve fungi were tested for antifungal activity including: Candida albicans ATCC 90028, Candida glabrata ATCC 66032, Candida parapsilosis ATCC 22019, Candida tropicalis ATCC 9928, Fusarium solani ATCC 36031, Fusarium moniliforme JLCC 31463, Fusarium oxysporum JLCC 30866, Aspergillus flavus IFM 55648, Aspergillus fumigatus IFM 40808, Aspergillus terrus JLCC 30844, Sporothrix schenckii JLCC 32757, and Cryptococcus neoformans ATCC 36556 (American Type Culture Collection, ATCC; Culture Collection of Jilin University, Mycology Research Center, JLCC; Institute for Food Microbiology, Medical Mycology Research Center, Chiba University, IFM). Cervical cancer cells (HeLa, ATCC CCL-2) and breast cancer cells (MCF-7, ATCC CRL-3435) were purchased from ATCC. Sabouraud medium was purchased from BD (Becton, Dickinson and company); the silver nitrate (AgNO3) used in this study was purchased from Sigma-Aldrich. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), RPMI 1640, and fetal bovine serum (FBS) were purchased from Invitrogen.
10
HeLa Cell Culture Protocol
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The HeLa CCL2 cell line was obtained from the ATCC collection (atcc.org , HeLa (ATCC® CCL-2™, RRID CVCL_0030) and cultured in modified complete Dulbecco’s modified Eagle’s medium (DMEM) at 37°C and 5% CO2. The sex of the cells is female. No additional cell line authentication was performed.
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