Normal nasopharyngeal epithelial cells NP69 (BNCC338439) and NPC cell line C666–1 (BNCC337872) were purchased from BeNa Culture Collection (BNCC, Beijing, China) in February, 2018. NP69 and C666–1 cells are not misidentification and contamination of human cell lines (ExPASy: SIB Bioinformatics Resource Portal, https://www.expasy.org/ ). These cells were seeded in culture solution (90%RPMI-1640 + 10%FBS) at 37 °C in a humid atmosphere with 5% CO2.
C666 1
Manufactured by BNCC
Sourced in China
The C666-1 is a laboratory centrifuge designed for general-purpose applications. It has a maximum speed of 6,000 rpm and can accommodate a range of rotor types to suit various sample volumes and tube sizes. The centrifuge features a digital display for speed and time control, as well as safety features like a lid lock and automatic power-off function.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Hone1, by BNCC (1 mentions)
Sune1, by Thermo Fisher Scientific (1 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
C666 1, by Thermo Fisher Scientific (1 mentions)
Keratinocyte serum free medium, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
5 protocols using c666 1
1
Culturing normal and cancer nasopharyngeal cells
2
Culturing Human Nasopharyngeal Carcinoma Cells
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The human NPC cell lines 5–8F, 6–10B and C666-1 were obtained from BeNa Culture Collection, Jiangsu, China. CNE2 and HONE1 were obtained from College of Pharmacy, Guilin Medical University, Guangxi, China. CNE1 was obtained from School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, China. Cells were maintained in DMEM or RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin. The cells were cultured at 37°C, in a humidified incubator with 95% O2 and 5% CO2 (Cheng et al., 2019 (link); Liang et al., 2019 (link)).
3
Evaluating ZFAS1 in Nasopharyngeal Carcinoma
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The human nasopharyngeal epithelial cell line NP69 and NPC cell lines (HONE1, CNE, HNE1 and C666‐1) were commercially obtained from BeNa Culture Collection (Beijing, China). NP69 cells were maintained in serum‐free keratinocyte medium containing 0.2 ng/mL human recombinant epidermal growth factor, 2% heat‐inactivated foetal bovine serum (FBS), 40 μg/mL bovine pituitary extract and 1% streptomycin and penicillin (Invitrogen, Carlsbad, CA, USA). CNE, HNE1 and C666‐1 cells were maintained in 90% RPMI‐1640 medium containing 10% FBS, and 1% streptomycin and penicillin. HONE1 cells were maintained in DMEM medium containing 5% FBS, 5% newborn calf serum and 1% streptomycin and penicillin. After incubation for 36 hours, cells were harvested to extract total RNA, and then real‐time RT‐PCR (qRT‐PCR) was carried out as mentioned below. Due to the highest expression level of ZFAS1 compared with other NPC cells, the HONE1 cell line was selected for the subsequent cell‐based experiments.
4
Culturing Human Nasopharyngeal Cell Lines
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The human NPC cell lines 6–10B and SUNE1 were obtained from Shanghai Biotech Bingsui Co. Ltd. (Shanghai, China). The human NPC cell line C666-1 and the normal nasopharyngeal cell line NP69 were purchased from BeNa Culture Collection (BNCC, China). The cell lines 6–10B, SUNE1 and C666-1cells were maintained at 37 °C in RPMI-1640 medium, 5–8F cells in DMEM medium, and NP69 in keratinocyte serum-free medium (Invitrogen, Carlsbad CA USA) in a 5% CO2 incubator.
5
Analyzing lncRNA ZFAS1, miR-7-5p, and ENO2 in NPC cell lines
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The human NPC cell lines SUNE-1, 5-8F, and C666-1 as well as normal nasal mucosal epithelial cell NP-69 were all bought from BeNa Culture Collection (Beijing, China). SUNE-1, 5-8F, and C666-1 were cultured in RPMI-1640 with 10% FBS.
RNA was detected by real-time quanti cation PCR.
Total RNA was disintegrated by Trizol (DP501, Tiangen Biochemical, China). The RNA was reverse transcribed into cDNA using a cDNA synthesis kit (KR211, Tiangen Biochemical, China) after checking the purity of RNA. Next, the expression of lncRNA ZFAS1, miR-7-5p and ENO2 mRNA were analyzed by ABI 7500 using a SYBR Green PCR kit (FP411, Tiangen Biochemical, China). U6 acted as a reference gene for miR-7-5p while GAPDH acted as a reference gene for lncRNA ZFAS1 and ENO2 mRNA.
RNA was detected by real-time quanti cation PCR.
Total RNA was disintegrated by Trizol (DP501, Tiangen Biochemical, China). The RNA was reverse transcribed into cDNA using a cDNA synthesis kit (KR211, Tiangen Biochemical, China) after checking the purity of RNA. Next, the expression of lncRNA ZFAS1, miR-7-5p and ENO2 mRNA were analyzed by ABI 7500 using a SYBR Green PCR kit (FP411, Tiangen Biochemical, China). U6 acted as a reference gene for miR-7-5p while GAPDH acted as a reference gene for lncRNA ZFAS1 and ENO2 mRNA.
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