Prmi 1640 medium

Manufactured by Thermo Fisher Scientific

Sourced in United States

The PRMI-1640 medium is a cell culture medium designed for the in vitro cultivation of a variety of cell types. It provides the necessary nutrients and growth factors to support the growth and maintenance of cells. The composition of the medium is optimized to maintain the physiological conditions required for cell proliferation and viability.

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Lab products found in correlation

45 protocols using prmi 1640 medium

Synthesis of Multifunctional Lipid Conjugates

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Polyethylene glycol (PEG) 2000-hydrazone-C18 (PHC),
O-stearoyl mannose (M-C18), and 4-(N)-stearoyl gemcitabine
(GemC18) were synthesized as previously described (27 (link), 29 (link), 30 (link)). Nile Red, bicinchoninic acid (BCA)
kit, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were
from Sigma-Aldrich (St. Louis, MO). D-(+)-mannose was from TCI America
(Portland, OR). Dimethyl sulfoxide (DMSO), analytical grade tetrahydrofuran
(THF) and methanol were from Thermo Fisher Scientific (Waltham, MA, USA). Mouse
macrophages (J774A.1), lung (LLC and TC-1), melanoma (B16–F10), and
pancreatic (Panc-02) tumor cell lines were from the American Type Culture
Collection (Manassas, VA). The M-Wnt mouse mammary tumor cells were generously
provided by Dr. Stephen D. Hursting at the University of North Carolina, Chapel
Hill. All cells were maintained in DMEM medium except the M-Wnt and TC-1 cells,
which were cultured in PRMI1640 medium (Invitrogen, Carlsbad, CA). The media
were supplemented with 10% fetal bovine serum (FBS), 100 U/mL of penicillin, 100
μg/mL of streptomycin, all from Invitrogen.

Alzhrani R.F., Xu H., Valdes S.A., Naguib Y.W, & Cui Z. (2021). Effect of surface mannosylation on the cytotoxicity and cellular uptake of stearoyl gemcitabine-incorporated, acid-sensitive micelles. Colloid and interface science communications, 43, 100441.

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Culturing immortalized placenta cells

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The HTR8/SVneo (HTR8) cell line was derived from a human first-trimester placenta explant immortalized by SV40 large T antigen [30 (link)]. The cells were cultured at 37°C with a 5% CO₂ atmosphere in phenol red-free PRMI 1640 medium (Gibco, Invitrogen, NY, USA) containing 10% fetal bovine serum (FBS), 100 IU/mL penicillin, and 100 µg/mL streptomycin. For steroid starvation, cells were cultured in phenol-free PRMI 1640 containing 5% charcoal-stripped FBS (Sigma, St. Louis, MO, USA).

Hu X.L., Shi S., Hou N.N., Meng Y., Li M., Liu A.X., Lu Y.C., Li J.Y., Sheng J.Z., Zhu Y.M, & Huang H.F. (2021). High Maternal Serum Estradiol in First Trimester of Multiple Pregnancy Contributes to Small for Gestational Age via DNMT1-Mediated CDKN1C Upregulation. Reproductive Sciences, 29(4), 1368-1378.

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Gastric Cancer Cell Line Transfection

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The human gastric epithelial cell line (GES-1 cells), human GC cell lines (BGC823 and SGC7901 cells), and human DDP-resistant GC cell lines (BGC823/DDP and SGC7901/DDP cells) were provided by the American Type Culture Collection (ATCC; Manassas, Virginia, USA). The above-mentioned cell lines were cultured in PRMI-1640 medium containing 10% FBS (Invitrogen, Carlsbad, CA, US) at 37°C with 5% CO₂. Small interfering RNAs (siRNAs) against LINC01857 (si-LINC01857#1: 5′-ATCTAACAGTTTCCGCTATTTTG-3′, si-LINC01857#2: 5′-AAGAGATAGAGGAAAAATGGATT-3′, and si-LINC01857#3: 5′-AGGAAAAATGGATTCAACAAATG-3′) and the negative control (si-NC) were provided by GenePharma (Shanghai, China). Cell transfection was performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA).

Chen Y, & Wu Y. (2022). Downregulation of LINC01857 Increases Sensitivity of Gastric Carcinoma Cells to Cisplatin. Computational and Mathematical Methods in Medicine, 2022, 3325686.

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Polarizing Prostate Cancer Macrophages

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Human prostate cancer cell line PC3 and RAW246.7 macrophages were purchased from BeNa Culture Collection (Beijing, China). PC3 cells and RAW246.7 macrophages were cultured in PRMI 1640 medium (Invitrogen, Gaithersburg, MD, USA) containing 10% fetal bovine serum (Invitrogen, USA) under the condition of 37°C and 5% CO₂. IL-4 (10 ng/mL) was used to stimulate RAW246.7 macrophages to obtain anti-inflammatory macrophages (M2),28 (link) while control macrophages (M0) were cultured in medium alone. Pro-inflammatory macrophages (M1) were induced by treating RAW246.7 macrophages with 50 ng/mL interferon (IFN)-γ and 10 ng/mL lipopolysaccharide (LPS).29 (link)

Xie W., Guo H., Zhang J., Hu L., Wu Y, & Wang X. (2021). Comprehensive Analysis of the Relationship Between Metabolic Reprogramming and Immune Function in Prostate Cancer. OncoTargets and therapy, 14, 3251-3266.

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Investigating CircRNA-miRNA-mRNA Axis in Multiple Myeloma

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Three MM cell lines (U266, MM1.S and NCI-H929) and human normal plasma cells (nPCs) were acquired from American Type Culture Collection (ATCC, Manassas, VA, USA). MM cells were seeded in PRMI-1640 medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% FBS (Invitrogen) with 5% CO₂ at 37 °C. Small interfering RNA against circ_0005615 (si-circ_0005615), miR-331-3p mimic (miR-331-3p), IGF1R overexpression vector (IGF1R) and their matched controls were harvested from Genechem (Shanghai, China). The oligo sequences are as follows: si-NC (5′-UCAUACGAACGAGAGAGGAA-3′), si-circ_0005615 (5′-ACCCCUAUAUUUCGAUCUUGA-3′), miR-NC(5′-CGAUCGCAUCAGCAUCGAUUGC-3′), miR-331-3p (5′-GCCCCUGGGCCUAUCCUAGAA-3′), anti-miR-NC (5′-CUAACGCAUGCACAGUCGUACG-3′) and anti-miR-331-3p (5′-UUCUAGGAUAGGCCCAGGGGC-3′). MM1.S and NCI-H929 cells were transfected with 30 nM oligo or 600 ng vector. The transfection experiment was carried out via applying Lipofectamine™3000 Kit (Invitrogen).

Zhang Q., Duan H., Yang W., Liu H., Tao X, & Zhang Y. (2023). Circ_0005615 restrains the progression of multiple myeloma through modulating miR-331-3p and IGF1R regulatory cascade. Journal of Orthopaedic Surgery and Research, 18, 356.

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Colorectal Cancer Specimen Collection and Cell Lines

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All CRC specimens and matched adjacent non-tumour tissues were obtained with informed consent from Nanfang Hospital, Southern Medical University (Guangzhou, China). Freshly frozen tumor samples from 35 CRC patients were selected for real-time PCR. 164 paraffin-embedded CRC samples were used for immunohistochemistry (IHC) and 150 samples for in situ hybridization (ISH). Complete follow-up, ranging from 1 to 117 months, and no patient received any pre-operative chemotherapy and radiotherapy.
The human CRC cell lines SW620, SW480, DLD-1, HCT116, and LoVo, normal colon epithelial cell line FHC, and the human embryonic kidney 293 T cell lines were obtained from American Type Culture Collection. M5 cell line, a subclone with enhanced metastasis ability, was derived from SW480 through in vivo selection in our laboratory. All of the CRC and FHC cell lines were cultured in PRMI 1640 medium (Gibco, USA), HEK-293 T was cultured in Dulbecco’s modified Eagle’s medium (DMEM; Gibco, USA). All the medium was added with 10% FBS (Gibco, USA). All the cells were cultured at 37 °C with 5% CO₂.

Yang M.H., Zhao L., Wang L., Ou-Yang W., Hu S.S., Li W.L., Ai M.L., Wang Y.Q., Han Y., Li T.T., Ding Y.Q, & Wang S. (2019). Nuclear lncRNA HOXD-AS1 suppresses colorectal carcinoma growth and metastasis via inhibiting HOXD3-induced integrin β3 transcriptional activating and MAPK/AKT signalling. Molecular Cancer, 18, 31.

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Cytotoxicity of Fusarium Polysaccharides

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The cytotoxicity effect of extracted polysaccharide of Fusarium was evaluated by MTT assay (Mosmann, 1983 (link)). Two human cancer cell lines were employed including HeLa cells and LCLs. These two cell lines were obtained from the cell culture unit of molecular biology laboratory, Karaj Islamic Azad University, Alborz, Iran.
The cells were grown in 96 well plate (Greiner, Germany) in PRMI 1640 medium (GIBCO, Grand Island, NY) supplemented with 10% fetal bovine serum. The cell suspension (10⁵ cell/ml) was seeded in every well and incubated at 37 °C for 48 h in 5% CO₂ for the formation of confluent monolayer. The monolayer of cells was exposed to various dilutions (10, 50, 100 mg/ml) of a polysaccharide extract for 24 h. The cell viability was measured using the MTT assay. The MTT solution (5 mg/ml) was added to the culture wells and the plates were incubated at 37 °C for 3 h. The optical density (OD) of the well contents was measured at 553 nm with an ELISA plate reader (Voller et al., 1978 (link)). ELISA is a simple method which enjoys some advantages such as portability of the equipment, hand-holding validation, and reliability for the analysis of samples (Kamkar et al., 2014 , Mozaffari Nejad et al., 2013 , Mozaffari Nejad et al., 2014 , Eslami et al., 2015 ). Cell control was maintained throughout the experiment and the assay was performed in triplicates.

Salehi B., Bayat M., Dezfulian M., Sabokbar A, & Tabaraie B. (2016). The assessment of anti-tumoral activity of polysaccharide extracted from terrestrial filamentous fungus. Saudi Journal of Biological Sciences, 25(6), 1236-1241.

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Modulation of miR-548c and Twist in EC and OC Cells

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The human EC cell lines (RL95-2 and HEC-1) and human OC cell lines (SKOV-3 and OVCAR3) were obtained from Chinese Academy of Sciences Committee on Type Culture Collection Cell Bank, Shanghai, China. These cells were cultured in DMEM/F12 medium (Invitrogen, Shanghai) or PRMI-1640 medium (Gibco, Carlsbad, CA, USA) supplemented with 10 % fetal bovine serum (FBS, Invitrogen, Shanghai). MiR-548c mimic (40 nM), antisense miR-548c inhibitor (40 nM), Twist siRNA (5 nM) and respective negative controls were purchased from Ambion (TX, USA), and were transfected using Lipofectamine 2000 (Invitrogen, CA, USA), according to the manufacturer’s instructions. Transient transfection of Twist cDNA plasmids (OriGene, MD, USA) were performed with Lipofectamine Plus reagent (Invitrogen, CA, USA) according to the manufacturer’s protocol.

Sun X., Cui M., Zhang A., Tong L., Wang K., Li K., Wang X., Sun Z, & Zhang H. (2016). MiR-548c impairs migration and invasion of endometrial and ovarian cancer cells via downregulation of Twist. Journal of Experimental & Clinical Cancer Research : CR, 35, 10.

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Molecular mechanisms of FKBP4 in cancer

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The cell lines PC9, H1975, and BEAS-2B were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China) and authenticated using STR profiling. The cells were cultured in PRMI-1640 medium (Gibco, MA, USA) supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin−streptomycin (Solarbio, Beijing, China) at 37 °C in a humidified atmosphere with 5% CO₂.
The small-interfering RNA (siRNA) targeting FKBP4 was synthesized by GenePharma (Shanghai, China). The following plasmids were purchased from BioSune (Shanghai, China): samples of plasmids encoding human Hsp90, IKKβ, and RelA; hemagglutinin (HA)-tagged IKKα, IKKβ, IKKγ, and Hsp90; Flag-tagged FKBP4 full-length (Flag-FKBP4-WT); Flag-tagged FKBP4-deleted PPIase domains (Flag-FKBP4ΔPPIase; 17-253 AA-truncated mutant); Flag-tagged FKBP4-deleted TPR domains (Flag-FKBP4ΔTPR; 269-386 AA-truncated mutant); Flag-tagged FKBP4 with a point mutant in the TPR domain (K354A). All transfections were performed with Lipofectamine 3000 (Invitrogen, MA, USA) according to the manufacturer’s instructions.
Lentiviruses carrying the FKBP4−RNA interference sequence (shR-FKBP4) or FKBP4-overexpression sequence were purchased from GeneChem (Shanghai, China). The stably transfected cells were further selected with puromycin (2 μg/ml) and validated by western blot.

Zong S., Jiao Y., Liu X., Mu W., Yuan X., Qu Y., Xia Y., Liu S., Sun H., Wang L., Cui B., Liu X., Li P, & Zhao Y. (2021). FKBP4 integrates FKBP4/Hsp90/IKK with FKBP4/Hsp70/RelA complex to promote lung adenocarcinoma progression via IKK/NF-κB signaling. Cell Death & Disease, 12(6), 602.

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Culturing Bladder Cancer Cell Lines

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The human bladder cancer cell lines T24 and 253J were purchased from the American Type Culture Collection (Manassas, VA, USA). The two cell lines, T24 and 253J, were cultured in PRMI-1640 medium (Gibco, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco, Australia). The cells were maintained at 37 °C in a humidified incubator with a constant air flow of 5% CO2.

Zhang C., Chang X., Chen D., Yang F., Li Z., Li D., Yu N., Yan L., Liu H, & Xu Z. (2019). Downregulation of HDGF inhibits the tumorigenesis of bladder cancer cells by inactivating the PI3K-AKT signaling pathway. Cancer Management and Research, 11, 7909-7923.

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