A novel source of viable peripheral blood mononuclear cells from leukoreduction system chambers

Abstract

LRSCs are a novel source of fully functional PBMNCs that can replace the more traditional sources of research-grade cellular products.

Tumor necrosis factor receptor-associated factor-1 enhances proinflammatory TNF receptor-2 signaling and modifies TNFR1–TNFR2 cooperation

“…Isolation and activation of primary T cells Peripheral blood mononuclear cells were isolated by Ficoll (PAA, Pasching) density-gradient centrifugation from cells retained in leukoreduction system chambers (Dietz et al, 2006). T cells were isolated with a MACS Seperator using CD3-specific beads (Milteny Biotech, Auburn, CA, USA).…”

High-density preculture of PBMCs restores defective sensitivity of circulating CD8 T cells to virus- and tumor-derived antigens

“…Human PBMCs were prepared from healthy donors from leukoreduction system chambers 16 or as heparinized venous blood from patients who had undergone myeloablation and allogeneic hematopoietic stem cell transplantation (HSCT) and were in complete hematologic remission. PBMCs were obtained by density gradient centrifugation and kept on ice until use within 1 hour, or freezing in 40% RPMI 1640 (Gibco), 50% AB-positive heat-inactivated human serum, and 10% dimethylsulfoxide (Sigma-Aldrich).…”

Detection of Constant Domain of Human T Cell Antigen Receptor Alpha-Chain Via Novel Monoclonal Antibody 7F18

“…(23) RPMI-cultured cells were incubated in 5% CO 2 atmosphere, while DMEM-cultured cells were incubated in 10% CO 2 at 37°C. Human peripheral blood mononuclear cells (PBMC) were Ficoll-purified from leukoreduction system chambers as described, (24) in accordance with Mayo Clinic's Institutional Review Board regulations. .…”

“…(26) For GST-Nck-SH3.1 pull-down assays, published methods were used. (24)(25)(26)(27) Briefly, lysate from JRT3.huCD8ab.LC13 cells was incubated on ice for 30 min in the presence of 10 mg/ mL of either APA1/1 (anti-CD3-epsilon, intracellular tail) or OKT3 (anti-human CD3-epsilon extracellular domain), followed by incubation with GST-Nck-SH3.1 beads overnight at 4°C. Captured TCR/CD3 complexes were subjected to reducing SDS-PAGE and subsequent Western blot analysis with 7F18.…”