Memory B cells were differentiated in vitro into plasmablasts and plasma cells, following the protocol described in Jourdan et al. (24 (link)). Briefly, CD27+ Memory B cells were first purified as bulk B cells by negative selection from PBMCs by magnetic isolation followed by positive selection for CD27+ cells (Stemcell Technologies, UK) following manufacture's protocol. Then seeded at1.5 × 105 cells/ml, and cultured for 4 days in the presence of CpG oligodeoxynucleotide 2006 (10 μg/ml; Invitrogen), soluble human his-rCD40L (50ng/ml; R&D Systems, UK) and anti-poly-his (5μ g/ml; R&D Systems, UK), IL-2 (20 U/ml; R&D Systems, UK), IL-10 (50 ng/ml; Miltenyi Biotec, UK) and IL-15 (10 ng/ml; Miltenyi Biotec, UK). At day 4 of culture, the cells were washed and cultured at 2.5 × 105 cells/ml with IL-2 (20 U/ml, Milteny Biotec, UK), IL-10 (50 ng/ml), IL-15 (10 ng/m, PreproTech, UK) and IL-6 (50ng/ml; Miltenyi Biotec, UK). At day 7 of culture, cells were washed and seeded with IL-6 (50 ng/ml, PreproTech, UK), IL-15 (10 ng/ml, PreproTech, UK) and IFN-α (500 U/ml; R&D Systems) for 3 days.
Cd27 cells
Manufactured by STEMCELL
Sourced in United Kingdom
CD27+ cells are a type of lymphocyte that express the CD27 surface marker. They play a role in the adaptive immune response and are involved in the differentiation and survival of B cells and T cells. The core function of CD27+ cells is to facilitate immune system signaling and regulation, though their precise applications may vary depending on the specific research or clinical context.
Automatically generated - may contain errors
Lab products found in correlation
Soluble human his rcd40l, by R&D Systems (2 mentions)
Anti poly his, by R&D Systems (2 mentions)
Cpg oligodeoxynucleotide 2006, by Thermo Fisher Scientific (2 mentions)
Il 10, by Miltenyi Biotec (2 mentions)
Il 10, by Thermo Fisher Scientific (2 mentions)
Il 15, by Thermo Fisher Scientific (2 mentions)
Interleukin 6 (il 6), by Miltenyi Biotec (2 mentions)
Interleukin 6 (il 6), by Thermo Fisher Scientific (2 mentions)
Ifn α, by R&D Systems (2 mentions)
Interleukin 2 (il 2), by Miltenyi Biotec (2 mentions)
Il 15, by Miltenyi Biotec (2 mentions)
Interleukin 2 (il 2), by R&D Systems (2 mentions)
2 protocols using cd27 cells
1
Differentiation of Memory B Cells into Antibody-Secreting Cells
2
In Vitro Differentiation of Memory B Cells
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Memory B cells were differentiated in vitro into plasmablasts and plasma cells, following the protocol described in Jourdan et al. (24 (link)). Briefly, CD27+ Memory B cells were first purified as bulk B cells by negative selection from PBMCs by magnetic isolation followed by positive selection for CD27+ cells (Stemcell Technologies) following the manufacturer’s protocol. The cells were then seeded at 1.5 × 105 cells/ml and cultured for 4 d in the presence of CpG oligodeoxynucleotide 2006 (10 μg/ml; Invitrogen, Paisley, U.K.), soluble human his-rCD40L (50 ng/ml; R&D Systems, Abingdon, U.K.) and anti–poly-his (5 μg/ml; R&D Systems), IL-2 (20 U/ml; R&D Systems), IL-10 (50 ng/ml; Miltenyi Biotec, Surrey, U.K.), and IL-15 (10 ng/ml; Miltenyi Biotec). At day 4 of culture, the cells were washed and cultured at 2.5 × 105 cells/ml with IL-2 (20 U/ml; Miltenyi Biotec), IL-10 (50 ng/ml), IL-15 (10 ng/ml; PeproTech, London, U.K.), and IL-6 (50 ng/ml; Miltenyi Biotec). At day 7 of culture, cells were washed and seeded with IL-6 (50 ng/ml; PeproTech), IL-15 (10 ng/ml; PeproTech), and IFN-α (500 U/ml; R&D Systems) for 3 d.
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