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Immunotoxin-Mediated Depletion of CD5+ T Cells from Bone Marrow for Graft-vs.-Host Disease Prophylaxis
Published in Adrian P. Gee, BONE MARROW PROCESSING and PURGING, 2020
Joseph H. Antin, Howard J. Weinstein, Cyril Bouloux, Barbara E. Bierer
The cells were treated in a mixture containing: RPMI-1640 with L-glutamine without sodium bicarbonate to which 5% HSA had been added.Ammonium chloride (5 mEq/ml injection, Abbott Laboratories, N. Chicago, IL), 1:250 dilution for treatment, of a final concentration of 20 mM.THAM solution (Tromethamine) (Abbott Laboratories, N. Chicago, IL, each 100 ml containing tromethamine 3.6 g or 30 mEq), to a final concentration of 5%, adjusted to pH 7.8 ± 0.15 at room temperature.ST1-IT (Sanofi Research, Montpellier, France) reconstituted in 5 ml 25% human serum albumin (HSA); 10−6M ricin A chain initial concentration, 1% or 10−8M ricin A chain final concentration.
Definition of HLA-Dw Determinants Using Homozygous Typing Cells and the Mixed Lymphocyte Culture
Published in M. Kam, Jeffrey L. Bidwell, Handbook of HLA TYPING TECHNIQUES, 2020
Two types of media are required. One type is used throughout the procedures to dilute blood and wash cells, for which Hanks balanced salt solution (HBSS), RPMI, or sterile phosphate buffer saline (PBS) can be used. The other medium, designated culture medium, is used to resuspend the cells before setting up the cultures, and it requires an extra source of protein (normally human serum is used). The culture medium consists of RPMI 1640 supplemented with 2 mM L-glutamine, 50 U/ml penicillin, 50 μg/ml streptomycin plus 10% pooled human serum (PHS). The medium should be filtered through a 0.22-μ filter.
Host-Parasite Interactions With Macrophages In Culture
Published in Hans H. Gadebusch, Phagocytes and Cellular Immunity, 2020
Lee S. F. Soderberg, Morris Solotorovsky
The culture conditions that are most often used for macrophages have been heavily influenced by the culture system designed by Mishell and Dutton80 to demonstrate plaque-forming cells among mouse spleen cells. The culture medium generally consists of a balanced salt solution, such as Earle’s, Hank’s, or Tyrode’s, usually used in conjunction with a nutritional medium. Eagle’s minimal essential medium that was used by Mishell and Dutton80 is the nutritional medium reported most often in recent literature. Medium 199 and RPMI 1640 are also frequently used to culture macrophages. Other media that are used for macrophage culture include NCTC 109, Trowell’s T81, McCoy 5A, and Dulbecco’s.
Construction and application of microneedle-mediated photothermal therapy and immunotherapy combined anti-tumor drug delivery system
Published in Drug Delivery, 2023
Jiaqi Weng, Gensuo Zheng, Jiaoli Wen, Jing Yang, Qingliang Yang, Xi Zheng, Qinying Yan
Anti-PD-1 peptide (Sequence: (SNTSESF)2KFRVTQLAPKAQIKE-NH2 Branched peptide) (molecular weight 3261.55) was supplied by DGpeptides Co., Ltd. (China). Monoterminal methoxy polyethylene glycol mercapto polymer was purchased from Shanghai Macklin Reagent Co., Ltd. (China). Polymethyl vinyl ether maleic anhydride copolymer was purchased from Shanghai JuWei International Trade Co., Ltd. Polydimethylsiloxane (PDMS, Sylgard 184) was obtained from Dow Corning (USA). RPMI 1640 was provided by Thermo Fisher Scientific Co., Ltd. (China). FITC-anti-mouse CD11c, APC-anti-mouse CD80, and PE-anti-mouse CD86 were purchased from Biolegend (USA). Calcein/PI Cell Viability/Cytotoxicity Assay Kit was purchased from Beyotime Biotechnology Co., Ltd. (China). Other chemical solvents used were of reagent grade and used as directed.
Study on co-delivery of pemetrexed disodium and Bcl-2 siRNA by poly-γ-glutamic acid-modified cationic liposomes for the inhibition of NSCLC
Published in Drug Development and Industrial Pharmacy, 2023
Xiaoyu Huang, Ruonan Song, Xiao Wang, Kongfang He, Rumeng Shan, Fei Xie, Guihua Huang
Cholesterol was purchased from Sinopharm (Shanghai, China). Egg yolk lecithin was supplied by Tywei (Shanghai, China). Stearylamine and γ-PGA were provided by Macklin (Shanghai, China). The sequences of Bcl-2 siRNA were 5′-GGGAGAACAGGGUACGAUATT-3′ (forward) and 5′-UAUCGUACCCUGUUCUCCCTT-3′ (reverse), obtained from GenePharm (Shanghai, China). PMX was obtained from Meilunbio (Dalian, China). RNase A was purchased from Yuanyebio (Shanghai, China). RPMI-1640 and DMEM with high glucose medium were provided by Thermo Fisher Scientific (Waltham, USA). Fetal bovine serum (FBS) was purchased from Tianhangbio (Zhejiang, China). DAPI, 4% paraformaldehyde, PMSF, PVDF membrane, and leukocyte diluent were obtained from Soleibao (Beijing, China). Lipo transfection reagent, lipo-6000, cell cycle kit and apoptosis kit, Hoechst 33342 staining solution, hematoxylin, and eosin (HE) staining kit and tris buffered saline with Tween (TBST) were purchased from Beyotimebio (Shanghai, China). MTT was purchased from Sigma (Aldrich, USA). TRNzol Universal reagent was obtained from Tiangen Biotech (Beijing, China). HiFiScript cDNA Synthesis kit and UltraSYBY Mixture were purchased from ComWin Biotech (Beijing, China). RIPA lysate, bovine serum albumin, 5× SDS-PAGE protein loading buffer, pre-stained protein marker, SDS-PAGE gel rapid preparation kit, Bcl-2 antibody, HRP-labeled goat anti-rabbit IgG, ECL luminescent solution were purchased from Wanleibio (Shenyang, China). The beta tubulin antibody was obtained from Bioways (Shanghai, China). All other organic reagents were of analytical reagent grade or higher.
Polycaprolactone and polycaprolactone triol blends to obtain a stable liquid nanotechnological formulation: synthesis, characterization and in vitro – in vivo taste masking evaluation
Published in Drug Development and Industrial Pharmacy, 2021
Juliana Emanuelli, Viviane Pagnussat, Katherine Krieser, Julia Willig, Andréia Buffon, Luiz A. Kanis, Stanley Bilatto, Daniel Souza Correa, Thaís F. Maito, Sílvia S. Guterres, Adriana R. Pohlmann, Irene C . Külkamp-Guerreiro
The cytotoxicity test of the saquinavir nanocapsules and the control nanocapsules was evaluated on human T lymphocytes, using the cell viability Trypan Blue exclusion assay according to the protocol described in [30]. This test was carried out with the approval of the Research Ethics Committee (CAAE 40234114.6.0000.5347, No. 1.025.942). The extraction of lymphocytes from whole blood was performed with Histopaque® reagent. The cells with the final volume of 100 μL were cultured in sterile 96-well microplates containing RPMI-1640, with 2 g of sodium bicarbonate, 100 μg/ml of streptomycin and 100 μg/ml of amphotericin. Cell culture with RPMI-1640 was analyzed as a control. Nanoformulations containing saquinavir in concentrations of 0.5 and 1 μM or control nanoformulations were added directly to the plate in a 1:10 ratio. The plate was incubated in a CO2 oven at 37 °C for 24 h.