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Chapter 4: Monodisperse Polymer Particles in Immunoassays and Cell Separation
Published in Alan Rembaum, Zoltán A. Tökés, Micro spheres: Medical and Biological Applications, 2017
Kjell Nustad, Havard Danielsen, Albrecht Reith, Steinar Funderud, Tor Lea, Frode Vartdal, John Ugelstad
Peripheral blood mononuclear cells (PBM) were obtained by Isopaque Ficoll gradient centrifugation.33 In these experiments 106 PBM were rosetted with 2 mg particles corresponding to a particle/cell ratio of approximately 20. Aliquots of PBM (5 × 106 cells/mℓ) were incubated in HBSS-1% FCS with particles (A), (B), and (C), respectively in plastic tubes for 30 min at room temperature. The tubes were carefully rotated for 1 min at 10-min intervals. Before separation, small samples were taken from the particle-cell suspensions and the number of rosette-forming cells was counted (Table 1). Magnetic separation of cells was carried out by letting the particle-cell mixture flow (1 to 2 mℓ/min) through a 3-mm plastic tube attached to a V-shaped iron block lined with cobalt-samarium magnets. Effluent cells were centrifuged and the cell pellet was resuspended in RPMI 1640 and counted on a Coulter DN cell counter. Cells incubated with particles (A), (B), and (C), as well as untreated PBM, were assayed for rosette formation with 2-aminoethyl-isothiouronium bromide-treated sheep erythrocytes (EAET) and for 3H-thymidine incorporation after stimulation with phytohemagglutinin (PHA), pokeweed mitogen (PWM), purified protein derivative (PPD), and irradiated allogeneic PBM as described elsewhere.34 The main results of these experiments are shown in Table 1.
Scaffolds for tracheal regeneration
Published in Gilson Khang, Handbook of Intelligent Scaffolds for Tissue Engineering and Regenerative Medicine, 2017
Doh Young Lee, Seong Keun Kwon
Multipotent or adult stem cells, such as MSCs, hematopoietic stem cells (HSCs), or amniotic fluid stem cells, can differentiate into diverse cell types. These stem cells are usually obtained from the recipient’s tissues (e.g., bone marrow, peripheral blood, or fat tissue) with essentially no ethical concerns and can be applied without immunosuppression. These adult stem cells have already been used in many trachea-related and trachea-nonrelated clinical settings.29,30–31 Mononuclear cells (MNCs) have become clinically interesting because they include stem and progenitor cells, can be isolated from the bone marrow or the peripheral blood, and can be actively mobilized to increase the yield of isolated cells.
Lymphocytes and Pulsing Magnetic Fields
Published in Andrew A. Marino, Modern Bioelectricity, 2020
Ruggero Cadossi, Giovanni Emilia, Giovanni Ceccherelli, Guiseppe Torelli
Mononuclear cells (monocytes and lymphocytes) can be separated from the other cells present by centrifugation on Ficoll-Hypaque 1077 (Sigma) (used 1: 1 to the total volume of the diluted plasma). In this way the mononuclear cells are separated on the basis of their density.
Safety and bioactivity assessment of aqueous extract of Thai Henna (Lawsonia inermis Linn.) Leaf
Published in Journal of Toxicology and Environmental Health, Part A, 2021
Orawan Khantamat, Nahathai Dukaew, Jirarat Karinchai, Teera Chewonarin, Pornsiri Pitchakarn, Piya Temviriyanukul
The human immortalized keratinocyte HaCaT cell was kindly provided by Dr. Ariyaphong Wongnoppavich, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand. Murine macrophage RAW 264.7 cell line was purchased from the American Type Culture Collection (ATCC, USA). Cells were cultured in Dulbecco’s Modified Eagles Medium (DMEM) containing 10% heat-inactivated fetal bovine serum (FBS) and 1% penicillin/streptomycin. Peripheral blood mononuclear cells (PBMCs) were separated from heparinized blood obtained from healthy adult volunteers by density gradient centrifugation using Ficoll-hypaque. Cells were then cultured in RPMI-1640 medium supplemented with 10% FBS, 2 mM glutamine, and 1% penicillin/streptomycin. All these cells were maintained at 37°C in a humidified atmosphere containing 5% CO2. The 70–80% confluent cells were applied for further experiments.
Triethylene glycol dimethacrylate: adjuvant properties and effect on cytokine production
Published in Acta Biomaterialia Odontologica Scandinavica, 2018
Sara Alizadehgharib, Anna-Karin Östberg, Ulf Dahlgren
Fresh blood cells from eight healthy blood donors were obtained from Sahlgrenska University Hospital in Gothenburg, Sweden. Peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation using Ficoll-Paque Plus (GE Healthcare Bio-Sciences, Uppsala, Sweden). The cells were resuspended in PBS, centrifuged, and resuspended in Dulbecco’s Modified Eagle’s Medium (D-MEM) (Invitrogen, Lidingö, Sweden) that was supplemented with 5% heat-inactivated human AB serum (Sigma Chemical Co., St. Louis, MO), 100 U/mL of penicillin and 100 μg/mL of streptomycin (Invitrogen). Cell viability was determined by staining with 0.4% Trypan Blue (Sigma-Aldrich, Steinheim, Germany) and counted under a microscope using a Bürker chamber.
Effects of exercise training on immunological factors in kidney transplant recipients; a randomized controlled trial
Published in Research in Sports Medicine, 2022
Nazi Hemmati, Sohrab Kazemi, Narges Jamshidian-Tehrani, Jamshid Roozbeh, Maryam Koushkie Jahromi, Mohsen Salesi, Meghdad Abdollahpour-Alitappeh, Mohammad Hossein Karimi
Twenty-four hours after the last exercise session, blood samples were collected from the patients and serum was separated from the samples by centrifuging and kept in −20°C until analysis. Peripheral blood mononuclear cells (PBMCs) were immediately isolated from the blood samples using Ficoll (Greiner Bio-One Ltd, Stonehouse, UK).