Ca Ski cells (ATCC CRL-1550) were grown to 80% confluence in 10% fetal calf serum (FCS) Dulbecco’s Modified Eagle Medium (DMEM) in 24-well cell culture plates (37°C, 5%CO2). Lactobacillus isolates (4.2x106 CFUs) were added in triplicate and co-cultured for 3 hours at 37°C (5%CO2), after which unbound bacteria were gently removed by washing the cells three times with PBS. Cells and bound bacteria were detached using 0.1% Triton X-100, serially diluted, plated onto MRS agar plates, and incubated anaerobically at 37°C for 48 hours. The percentage of adhered bacteria was calculated as the number of bound cells compared to total number of bacterial cells initially added. Experiments were repeated three times.
Caski cell
Manufactured by American Type Culture Collection
Sourced in United States
CaSki cells are a human cervical epithelial cell line derived from a squamous cell carcinoma of the cervix. They are a well-characterized and widely used in vitro model for studying cervical cancer and human papillomavirus (HPV) infection.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Hela cell, by American Type Culture Collection (4 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Pen strep solution, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Cc7t vgh cells, by American Type Culture Collection (2 mentions)
Influx cell sorter, by BD (1 mentions)
Fetal bovine serum (fbs), by Gemini Bio (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Hk 2 cell, by American Type Culture Collection (1 mentions)
Keratinocyte sfm, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by American Type Culture Collection (1 mentions)
U2os cell, by American Type Culture Collection (1 mentions)
5 aza 2 deoxycytidine 5azadc, by Epigentek (1 mentions)
Fetal bovine serum (fbs), by Lonza (1 mentions)
Hela cell, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Penicillin and streptomycin, by Beyotime (1 mentions)
Temsirolimus, by Abcam (1 mentions)
18 protocols using caski cell
1
Lactobacillus Adhesion to Cervical Cancer Cells
2
Evaluating 225Ac-BAPC Uptake in CasKi Cells
3
Adoptive Transfer of Transgenic T Cells
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Pmel-1 (gp100) TCR and OT-I transgenic mice were obtained from the Jackson Laboratory (Bar Harbor, Maine, USA) and housed at the National Institutes of Health (Bethesda, Maryland, USA). Splenocytes from pmel-1, OT-I, and P14 transgenic mice were used as donors for adoptive cell therapy. All experiments were conducted with the approval of the National Cancer Institute Animal Care and Use Committee.
The B16 tumor cell line (Cat. No. CRL-6475, ATCC, Manassas, Virginia, USA) was used for in vivo experiments. The B16-OVA line was established by transducing B16 cells with a retrovirus expressing the mKate2-SIINFEKL fusion protein. Transduced cells were then sorted by mKate2 fluorescence using a BD influx cell sorter (BD Biosciences, San Jose, California, USA). The cell line SS4050 is an HPV-16+ human leucocyte antigen (HLA)-A*0201+ cervical cancer line that was generated in our laboratory from a lung metastasis. CaSki cells were purchased from ATCC (Cat. No. CRM-CRL-1550). The melanoma cell line 624 was generated previously by the Surgery Branch of the National Cancer Institute (Bethesda, Maryland, USA). All tumor cell lines were cultured using Dulbecco Modified Eagle Medium with 10% heat-inactivated fetal bovine serum (Gemini Bio-Products, Sacramento, California, USA) at 37°C in an atmosphere containing 5% CO2.
The B16 tumor cell line (Cat. No. CRL-6475, ATCC, Manassas, Virginia, USA) was used for in vivo experiments. The B16-OVA line was established by transducing B16 cells with a retrovirus expressing the mKate2-SIINFEKL fusion protein. Transduced cells were then sorted by mKate2 fluorescence using a BD influx cell sorter (BD Biosciences, San Jose, California, USA). The cell line SS4050 is an HPV-16+ human leucocyte antigen (HLA)-A*0201+ cervical cancer line that was generated in our laboratory from a lung metastasis. CaSki cells were purchased from ATCC (Cat. No. CRM-CRL-1550). The melanoma cell line 624 was generated previously by the Surgery Branch of the National Cancer Institute (Bethesda, Maryland, USA). All tumor cell lines were cultured using Dulbecco Modified Eagle Medium with 10% heat-inactivated fetal bovine serum (Gemini Bio-Products, Sacramento, California, USA) at 37°C in an atmosphere containing 5% CO2.
4
Studying Cellular O-GlcNAcylation and PIP3 Production
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CaSki cells (ATCC CRL-1550) were maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin at 37 °C in 5% CO2 (Life Technologies). cDNA transfections were performed with Lipofectamine 2000 reagent (Life Technologies). For the study of O-GlcNAcylation in different cell compartments by BRET, 3.6 × 105 cells/well were transfected with 0.5 µg cDNAs coding for cytosol-, nucleus- or plasma membrane-targeted BRET O-GlcNAc-biosensors22 (link). For the study of PIP3 production by BRET, 3.6 × 105 cells/well were transfected with 0.7 µg Luc-Akt-PH and 0.3 µg pYFP-Mem cDNAs16 ,30 .
5
Cell Culture Conditions Detailed
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All cells were cultured
at 37 °C in 5% CO2. CaSki cells (ATCC, CRL-1550) were
cultured in RPMI 1640 medium (ATCC, 30-2001) supplemented with 1 Pen
Strep solution (Gibco, 15140-122) and 10% fetal bovine serum (Sigma,
12003C). HK-2 cells (ATCC, CRL-2190) were cultured in Keratinocyte
SFM (Gibco, 17005042) supplemented with bovine pituitary extract and
human recombinant EGF.
at 37 °C in 5% CO2. CaSki cells (ATCC, CRL-1550) were
cultured in RPMI 1640 medium (ATCC, 30-2001) supplemented with 1 Pen
Strep solution (Gibco, 15140-122) and 10% fetal bovine serum (Sigma,
12003C). HK-2 cells (ATCC, CRL-2190) were cultured in Keratinocyte
SFM (Gibco, 17005042) supplemented with bovine pituitary extract and
human recombinant EGF.
6
Cell Culture of Cervical Cancer Lines
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CaSki cells (CRL-1550, ATCC, United States) were cultured in RPMI-1640 medium (SH30809.01B, Hyclone, Logan, UT, United States) with 10% fetal bovine serum (Ausbian, Austrilia), 100 U/ml penicillin, and 100 mg/ml streptomycin in a humidified incubator with 5% CO2 at 37°C. C-33A cells (TCHu176, Shanghai Cell Bank, Chinese Academy of Sciences, China) were cultured in Dulbecco’s modified Eagle medium (SH30022.01, Hyclone, Logan, UT, United States) containing 10% fetal bovine serum at 37°C with 5% CO2.
7
Epigenetic Regulation in Cancer Cell Lines
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Ca Ski cells (ATCC, Manassas, VA, USA) were maintained in RPMI-1640 medium while SiHa, C33-A and U2OS cells (ATCC) were maintained in Dulbecco's modified Eagle medium (DMEM), both supplemented with 10% (v/v) fetal bovine serum (Lonza, Basel, Switzerland) in a humidified CO2 incubator at 37°C. Ca Ski and SiHa cells, two HPV16 cell lines, were treated with 5-aza-2′-deoxycytidine (5azadC) (Epigentek, Farmingdale, NY, USA) at a final concentration of 0.25 μM for 48h or 96h. The treatment medium was renewed every day.
8
Culturing Human Cervical Cancer Cells
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Human cervical adenocarcinoma HeLa cells and human cervical squamous cell carcinoma CaSki cells were provided from ATCC (Manassas, VA, USA). HeLa cells were grown in Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Grand Island, NY, USA) with 10% fetal bovine serum (FBS; Gibco, Grand Island, NY, USA) and 1% penicillin–streptomycin (Pen–Strep) solution (Invitrogen, Carlsbad, CA, USA). A Roswell Park Memorial Institute 1640 (RPMI-1640) medium with 10% FBS and 1% Pen-Strep solution was used to maintain CaSki cells. All cell lines were incubated at 37 °C in a humidified atmosphere with 5% CO2.
9
Cell Culture of Cervical Epithelial Lines
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Normal cervical epithelial cell line Ect1/E6E7 cells (ATCC, Manassas, VA, USA) were cultured in a Roswell Park Memorial Institute (RPMI)-1640 liquid culture medium (ThermoFisher Scientific, Shanghai, China) and CC cell lines C-33A (bio-69458), HeLa (bio-68123), SiHa (bio-69163), and CaSki cells (bio-69460) (ATCC, Manassas, VA, USA) were cultured in a Dulbecco's modified Eagle medium (DMEM)-F12 liquid culture medium (Ther-moFisher Scientific, Shanghai, China), with a temperature of 37℃ and an air condition of 5% CO 2 . Both cell media were added with 10% fetal bovine serum (ThermoFisher Scientific, Shanghai, China) and 100 U/mL penicillin and streptomycin (Beyotime, Shanghai, China) in advance.
10
Cervical Cancer Cell Line Cultivation
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Cervical cancer cell lines HeLa cells (BCRC#60005, Hsinchu, Taiwan) and CC7T/VGH cells (BCRC#60195), and CaSki cells (ATCC#CRL-1550) were used and cultured in DMEM supplemented with 10% fetal bovine serum (ThemoFisher Scientific) in a humidified atmosphere of 95% air and 5% CO2 at 37 °C. The UBE2C inhibitor CCI779 (Temsirolimus) was from Abcam.
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