Non-cancerous, immortalized human lung bronchial epithelial cell line BEAS-2B was purchased from Zhong Qiao Xin Zhou Biotechnology (Shanghai, China). Normal fetal lung fibroblast cell HFL1; human LUAD cell lines Calu-3, NCI-H1975, and NCI-H1395; LUSC line NCI-H520; and other NSCLC cell lines A549 and NCI-H1299 were purchased from Procell (Wuhan, China). LUSC lines NCI-H226 and SK-MES-1 were purchased from Shanghai Chinese Academy of Science Cell Bank (Shanghai, China). BEAS-2B was grown in Bronchial Epithelial Cell Growth Medium BulletKit (BEGM; Lonza, Basel, Switzerland). HFL1 was cultured in Ham’s F-12K medium (Gibco, Pittsburgh, PA, USA). Calu-3 and SK-MES-1 were cultured in Eagle’s minimum essential medium (MEM; Gibco). NCI-H1975, NCI-H1395, NCI-H1299, NCI-H520, and NCI-H226 were maintained in RPMI-1640 (Gibco). All cell lines, except for BEAS-2B, were supplemented with 10% fetal bovine serum (FBS), streptomycin (100 μg/mL), and penicillin G (100 U/mL; Gibco). All cell lines were authenticated by short tandem repeat (STR) profiling. Cells were incubated at 37°C in a humidified atmosphere with 5% CO2.
Nci h1975
Manufactured by Procell
Sourced in China
The NCI-H1975 is a human non-small cell lung cancer (NSCLC) cell line. It is a widely used model for in vitro studies in oncology research.
Automatically generated - may contain errors
Lab products found in correlation
Nci h1975, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Sk mes 1, by Thermo Fisher Scientific (2 mentions)
Nci h1299, by Thermo Fisher Scientific (2 mentions)
Nci h1299, by Procell (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Beas 2b, by Procell (2 mentions)
H1299, by Procell (2 mentions)
Ham s f 12k medium, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin g, by Thermo Fisher Scientific (1 mentions)
Beas 2b, by Thermo Fisher Scientific (1 mentions)
Nci h226, by Thermo Fisher Scientific (1 mentions)
Calu 3, by Thermo Fisher Scientific (1 mentions)
Nci h520, by Thermo Fisher Scientific (1 mentions)
Beas 2b, by Shanghai Zhong Qiao Xin Zhou Biotechnology (1 mentions)
Bronchial epithelial cell growth medium bullet kit begm, by Lonza (1 mentions)
Nci h520, by Procell (1 mentions)
Calu 3, by Procell (1 mentions)
9 protocols using nci h1975
1
Characterization of NSCLC Cell Lines
2
NSCLC Cell Line Knockdown Study
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The NSCLC cell lines NCI-H157, NCI-H1975, H1299 and A549, along with the normal lung epithelial cell line BEAS-2B, were obtained from Procell Life Science & Technology Co., Ltd. The identification method of these cell lines was short tandem repeat (STR) profiling. These cells were cultured using the following conditions: i) RPMI-1640 medium supplemented with 10% fetal bovine serum (both from Invitrogen; Thermo Fisher Scientific, Inc.); ii) incubation in a cell culture incubator at 5% CO2/37°C. The small interfering RNA (siRNA) that targeted SLCO4A1 was obtained from Suzhou GenePharma Co., Ltd. (Infection concentration: 5.5 μl per 100,000 cells.). The sequences of all siRNAs are included in Table SIV . Lipofectamine™ 3000 (Invitrogen; Thermo Fisher Scientific, Inc.) was used for transient transfection of these cell lines, following the manufacturer's instructions. After 48 h of transfection, qPCR and western blotting were performed to assess knockdown efficiency. Based on the results, suitable siRNA sequences were selected for further investigation.
3
EGFR Mutant Lung Cancer Cell Lines
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Two lung cancer cell lines containing L858R point mutation (NCI-H1975) and the wild-type variant (A549) of EGFR as the simulative positive and negative specimens, respectively, were purchased from Procell Life Science Co., Ltd. (Wuhan, China). The cell lines were cultured in the RPMI 1640 medium (HyClone Co., Ltd., Logan, UT, USA) with 10% fetal bovine serum (Gemini Bio Co., Ltd., West Sacramento, CA, USA) using a cell incubator (MCO-5AC, SANYO Electric Co., Ltd., Osaka, Japan) at 37 °C with 5% CO2. The cells at the logarithmic phase were detached using 0.25% trypsin-EDTA (Gibco Co., Ltd., Grand Island, NY, USA) for the subsequent study.
The cancerous tissues were surgically resected from 40 patients diagnosed with non-small cell lung cancer by the expert pathologist at the First Affiliated Hospital of Xi’an Jiaotong University (Xi’an, China) with informed consent. Then, all the cancerous tissues were formalin-fixed and paraffin-embedded for the subsequent study. This research was approved by the Ethics Committee of the College of Life Sciences, Northwest University (Xi’an, China), and all human-related protocols were performed in accordance with the Declaration of Helsinki.
The cancerous tissues were surgically resected from 40 patients diagnosed with non-small cell lung cancer by the expert pathologist at the First Affiliated Hospital of Xi’an Jiaotong University (Xi’an, China) with informed consent. Then, all the cancerous tissues were formalin-fixed and paraffin-embedded for the subsequent study. This research was approved by the Ethics Committee of the College of Life Sciences, Northwest University (Xi’an, China), and all human-related protocols were performed in accordance with the Declaration of Helsinki.
4
Investigating CHCHD4 Regulation in LUAD Cells
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Four human LUAD cell lines, NCI-H1975, H1299, A549, and H23, and the human bronchial epithelioid cell line 16-HBE, were supplied by Procell (Wuhan, China). All cell lines were maintained in RPMI-1640 medium (Hyclone) supplemented with 10% FBS (S660JJ, BasalMedia, Shanghai, China) and 1% streptomycin/penicillin (P1400, Solarbio, Beijing, China) in a humidified atmosphere of 5% CO2 at 37 °C. Small interfering RNAs against CHCHD4 (si-CHCHD4-1, si-CHCHD4-2, and si-CHCHD4-3) and their negative control (si-NC) were purchased from RiboBio (Guangzhou, China). Short hairpin RNAs (shRNAs) for CHCHD4 (sh-CHCHD4), and shRNA negative control (sh-NC) were obtained from Genechem (Shanghai, China). The pcDNA3.1-USF1 vector, pcDNA3.1-MYC vector and their empty vector were constructed by Tsingke Biotechnology Co., Ltd. Transfection was performed using Lipofectamine 3000 (Invitrogen, USA) according to the manufacturer's instructions.
5
Lung Adenocarcinoma Tissue and Cell Line Protocol
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This study includes five pairs of lung adenocarcinoma (LUAD) tissues from Space Central Hospital. The ethical approval for this study was from Space Central Hospital (20,200,511-JJJHZ-01). Written informed consents were obtained from each patient. Four NSCLC cell lines (A549, NCI-H1299, HCC827, and NCI-H1975) and human bronchial epithelial cell line HBEC were obtained from Procell Life Science&Technology Co.,Ltd (Wuhan, China). A549 cells were cultured in DMEM (High glucose, Gibco, Grand Island, NY, USA, 11,965–092) with 10% FBS (Gibco) and 1% penicillin/streptomycin (P/S, Procell). NCI-H1299, HCC827, and NCI-H1975 cells were cultured in RPMI-1640 medium (Gibco) with 10% FBS (Gibco) and 1% penicillin/streptomycin (P/S, Procell). HBEC cells were cultured in the bronchial epithelial growth medium (BEGM, Lonza, CC-3170). All the cells were cultured at 37 °C in a 5% CO2 incubator.
6
Overexpression of RNF182 and p65 in Lung Cancer Cells
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Human normal lung epithelial cells BEAS‐2B (CL‐0496), human LUAD cells A549 (CL‐0016) and NCI‐H1975 (CL‐0298), and 293T cells (CL‐0005) were procured from Procell Life Science & Technology. BEAS‐2B and 293T cells were incubated in Dulbecco's modified Eagle's medium, A549 cells in F12K medium, and NCI‐H1975 cells in Roswell Park Memorial Institute‐1640. All media were supplemented with 10% fetal bovine serum (FBS), 1% antibiotics, and 1% glutamine. The culture condition for all cells were 37°C with 5% CO2.
The overexpression plasmids of RNF182 and p65 (oe‐RNF182 and oe‐p65), the negative control (NC) plasmids, and the lentivirus vector for packaging were procured from VectorBuilder Inc. The lentivirus of oe‐RNF182 was puromycin‐resistant, while that of oe‐p65 was hygromycin‐resistant. The cells were treated with the lentivirus vectors until reaching 60% confluence. After 48 h,14 stably transfected cells were screened by the addition of corresponding antibiotics.
The overexpression plasmids of RNF182 and p65 (oe‐RNF182 and oe‐p65), the negative control (NC) plasmids, and the lentivirus vector for packaging were procured from VectorBuilder Inc. The lentivirus of oe‐RNF182 was puromycin‐resistant, while that of oe‐p65 was hygromycin‐resistant. The cells were treated with the lentivirus vectors until reaching 60% confluence. After 48 h,
7
Genomic DNA Extraction from Cell Lines and Whole Blood
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Whole blood was collected from healthy individuals and the genomic DNA was extracted as control for the development of PCR assays. Heterozygous control DNA was extracted from lung cancer cell lines, NCI-H-1650 and NCI-H-1975 (purchased from Procell, Wuhan, China), which contain the heterozygous delE746-A750 mutation in exon 19 and L858R mutation in exon 21, respectively. Wild-type DNA was extracted from the NCI-A549 cell line (purchased from Procell, Wuhan, China). The cell lines were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (GE Healthcare Hyclone, United Kingdom). All cells were maintained in 5% CO2 at 37 °C (Thermo Forma; America).
Genomic DNA from whole blood or cell lines was extracted using whole blood genomic DNA isolation kit from Xi’an GoldMag Nanobiotech Co., (Xi’an, Shanxi, China) according to the manufacturer’s instructions.
Genomic DNA from whole blood or cell lines was extracted using whole blood genomic DNA isolation kit from Xi’an GoldMag Nanobiotech Co., (Xi’an, Shanxi, China) according to the manufacturer’s instructions.
8
Lung Cancer Tissue and Cell Line Sourcing
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This research was approved by the Ethics Committee of The Fourth Hospital of Hebei Medical University. Five pairs of lung cancer tissues and adjacent tissues were derived from patients in the Fourth Hospital of Hebei Medical University with signed informed consent. The tissue samples were preserved at −80°C until subsequent detection. Lung cell lines A549, SK-MES-1, NCI-H1437, and NCI-H1975 were purchased from Procell Life Science&Technology Co., Ltd (Wuhan, China), and NCI-H2170 was purchased from Zhongqiaoxinzhou Biotech Co., Ltd (Shanghai, China). A549 was cultured in Ham’s F-12 K (PM150910, Procell Life Science) with 10% fetal bovine serum (FBS; SH30084.03, Hyclone, USA), while SK-MES-1 was maintained in MEM (41,500–067, Gibco, USA) with 10% FBS. NCI-H1437, NCI-H2170, and NCI-H1975 were grown in RPMI1640 (31,800–014, Gibco) with 10% FBS. All cells were cultured in an incubator (HF-90, Shanghai Lishen, China) of 95% humidity, 5% CO2 at 37°C.
9
Cell Line Cultivation Protocol
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The Beas-2B, A549, 95D, PC-9, and NCI-H1975 cell lines were purchased from ATCC (Manassas, USA). DMEM (Procell, Wuhan, China) supplemented with 10% fetal bovine serum (FBS; WISENT, Nanjing, China) and 1% penicillin/streptomycin was used to culture the Beas-2B, A549 and PC-9 cell lines. The 95D and NCI-H1975 cell lines were cultured in RPMI 1640 medium (Procell) supplemented with 10% FBS and 1% penicillin/streptomycin. Cells were incubated in a standard incubator with 5% CO
2 at 37°C.
2 at 37°C.
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