Whole blood was collected in heparinized tubes, diluted with PBS, and layered onto Ficoll-Hypaque (specific gravity, 1.086; Life Technologies, Grand Island, NY) for cell separation as previously described [6 (link)]. The isolated lymphocytes were resuspended in RPMI 1640 media supplemented with 10% human AB serum (Cambrex Biosciences, East Rutherford, NJ) in the presence of 5% CO2 at 37°C. The cell number and mean cell volume were determined using a Coulter channelyzer (Coulter Electronics, Hialeah, FL). The lymphocytes were suspended at a concentration of 1 x 107 cells/mL for all experiments and were used fresh.
Human ab serum
Manufactured by Cambrex
Sourced in Cameroon
Human AB serum is a component derived from human blood that contains a diverse array of proteins, growth factors, and other biomolecules. It serves as a versatile supplement for cell culture media, supporting the growth and maintenance of various cell types in laboratory settings.
Automatically generated - may contain errors
4 protocols using human ab serum
1
Lymphocyte Isolation from Whole Blood
2
Isolation of Peripheral Blood Cells from CLL Patients
3
PBMC Lymphocyte Culture Stimulation
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Lymphocyte culture was conducted as described previously.68 (link) Briefly, isolated PBMC were resuspended in the RPMI-1640 medium (Sigma) containing 20 mM HEPES, 2 mM L-glutamine, streptomycin (100 μg ml−1), penicillin (100 U ml−1), 50 μM 2-mercaptoethanol and 10% human AB serum (Cambrex Biosciences) and were cultured with the antigens. B19 VP2 VLPs were used at 1.50 and 0.5 μg ml−1, and the HBoV1 VLP and Candida albicans control antigens at 1.50 and 2.5 μg ml−1, respectively.
4
Isolation of PBMCs from CLL Patients
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We used two protocols to obtain peripheral blood samples from patients with CLL and from healthy donors. All participants signed an informed consent form, and the protocols were approved by The University of Texas MD Anderson Cancer Center institutional review board. Samples were collected in heparinized tubes and diluted in phosphate-buffered saline (PBS). Peripheral blood mononuclear cells (PBMCs) were isolated using the standard Ficoll-Hypaque (Life Technologies, Carlsbad, CA, USA) density gradient centrifugation, as described previously [7 (link)]. The isolated PBMCs were washed twice and then were suspended in Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with 10% human AB serum (Cambrex Biosciences, East Rutherford, NJ, USA) with 5% carbon dioxide at 37°C and maintained at 107 cells/mL. Cell number and cell size were determined by a Coulter channelyzer (Beckman Coulter; Brea, CA, USA).
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