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Human buffy coat

Cambridge Bioscience has a close collaboration with Research Donors, London-based clinic dedicated to the collection and processing of human blood for research purposes. esearch Donors is a HTA licenced, ISO 9001 2015 certified establishment with Research Ethics (REC) approval as a Research Tissue bank, as well as participating in the UK NEQAS QA scheme.

Our buffy coats are prepared according to each researcher's exact requirements, with delivery available on the same or next day after collection, offering access to human buffy coat whenever your research demands.

Buffy coat is prepared by the centrifugation of anticoagulated whole blood and it consists of a concentrated suspension of white blood cells and platelets. It is frequently used to isolate PBMCs and other specific white blood cell populations.
Buffy coat
Benefits of our human blood buffy coat service
• Same day delivery possible in south of UK or overnight delivery across Europe
• Specific donor/protocol requests accommodated
• Optional donor COVID-19 testing available
• Choice of anticoagulant and storage device
• Chilled or ambient delivery temperature
• Large donor pool
• Comprehensive donor information provided
• Serology screening prior to dispatch
• Samples fully consented including for genetic analysis and commercial research purposes

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Buffy coat sample validation data

Pre-freeze pheno buffy coat

Post-thaw pheno buffy coat

Left: Flow cytometry was used to assess the expression of CD3, TCR α/β TCR Vδ1, TCR Vδ2, CD14 and CD56 within PBMC freshly-isolated from buffy coats from three healthy donors. The data for αβT cells (CD3+TCR α/β +), Vδ1 yδT cells (CD3+TCR Vδ1+), Vδ2 yδT cells (CD3+TCR Vδ2+), monocytes (CD3-CD14+) and NK cells (CD3-CD56) is shown as mean percentage + SEM. Right: Flow cytometry was used to assess the expression of CD3, TCR α/β, TCR Vδ1, TCR Vδ2, CD14 and CD56 within PBMC immediately post cryorecovery in 10% DMSO from buffy coats from three healthy donors. The data for αβT cells (CD3+TCR α/β +), Vδ1 yδT cells (CD3+TCR Vδ1+), Vδ2 yδT cells (CD3+TCR Vδ2+), monocytes (CD3-CD14+) and NK cells (CD3-CD56) is shown as mean percentage + SEM.
Data courtesy of Jonathan Fisher et al, UCL Great Ormond Street Institute of Child Health, London, UK.

Post thaw viability & recovery buffy coat

PBMCs were cryopreserved in 10% DMSO and post-thaw viability was determined via trypan blue exclusion. Cell counts pre- and post-cryopreservation were used to calculate the percentage recovery. Mean percentage + SEM and individual values are shown for PBMCs from buffy coats from three healthy donors. Cell recovery after cryopreservation was >50%.
Data courtesy of Jonathan Fisher et al, UCL Great Ormond Street Institute of Child Health, London, UK.

Transduction impact on viability

Thawed T cell expandibility, transducibility and the impact of transduction on T cell viability” – Cryopreserved PBMC from buffy coats from three healthy donors were thawed, rested overnight in complete media (RPMI1640 supplemented with L- Glutamine and 10% FCS) and stimulated with a translationally relevant T cell expansion cocktail; specifically, PBMC received a single does of TCR stimulus post-thaw and were cultured in complete media containing 100 IU/mL IL-2. The expansion was supplemented with fresh IL-2 containing media every 2-3 days. T cell viability was assessed using flow cytometry after 12 days of stimulation for cell viability. Day 12 overall cell viability was consistently >60% for all samples, with marginal impact post-transduction. Mean percentage + SEM and individual values are shown for n=3.
Data courtesy of Jonathan Fisher et al, UCL Great Ormond Street Institute of Child Health, London, UK.

Transducibility

Thawed T cell transducibility” – Cryopreserved PBMC from three healthy donors were thawed, rested overnight in complete media (RPMI1640 supplemented with L- Glutamine and 10% FCS) and stimulated with a translationally relevant T cell expansion cocktail; specifically, PBMC received a single does of TCR stimulus post-thaw and were cultured in complete media containing 100 IU/mL IL-2. The expansion was supplemented with fresh IL-2 containing media every 2-3 days. T cell transducibility was assessed by viral transduction using a translationally-relevant lentiviral vector and protocol. T cell transduction efficiency was assessed on day 12 of expansion. T cells of buffy coats were higly and efficiently transducible with a lentiviralmultiplicity of infection of 4, with a mean efficiency of >90%. Data is shown for three donor buffy coat samples. Mean percentage + SEM and individual values are shown for n=3.
Data courtesy of Jonathan Fisher et al, UCL Great Ormond Street Institute of Child Health, London, UK.

Transduction impact on proliferation

Thawed T cell expandability, transducibility and the impact of transduction on T cell viability” – Cryopreserved PBMC from three healthy donors were thawed, rested overnight in complete media (RPMI1640 supplemented with L- Glutamine and 10% FCS) and stimulated with a translationally relevant T cell expansion cocktail; specifically, PBMC received a single does of TCR stimulus post-thaw and were cultured in complete media containing 100 IU/mL IL-2. The expansion was supplemented with fresh IL-2 containing media every 2-3 days. T cell expansion was assessed using flow cytometry and counting beads after 12 days of stimulation for cell viability. While non-transduced buffy coat T cells expanded at a mean >120-fold, transduced T cells expanded to a similar extent at a mean 100-fold. Mean percentage + SEM and individual values are shown for n=3.
Data courtesy of Jonathan Fisher et al, UCL Great Ormond Street Institute of Child Health, London, UK.

These products are for research use only and not for any diagnostic, therapeutic, human or animal use. It is your responsibility to ensure that you have necessary licensing and ethics approvals in place for the use and storage of any human tissue related material supplied by Cambridge Bioscience.