Flow Diagram of Immune Cell Preparation

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Flow Diagram of Immune Cell Preparation
Leukopack
UnPurified Leukopheresis Product
NK Cell
Elutriation
PBMC Prep via Ficoll Separation
Monocyte Prep
PBL
CD4 T Cell Purification via RosetteSep Kit
B Cell Purification via RosetteSep Kit
T Cell Purification via RosetteSep Kit
CD8 Purification via Miltenyi AutoMax
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PBMC Count
PBL
Mono cytes
PBMCs are isolated by subjecting the apheresis
product to a Ficoll gradient. PBLs and Monocytes
are separated by elutriation. Monocytes isolated
this way are very pure (gt95). If additional
purity is required, negative selection can be
performed for a additional cost.
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CD4 T Cell Purity
CD4 T cells are negatively selected from the
PBLs by using antibodies which target cell
surface antigens on human hematopoietic cells
(CD8, CD16, CD19, CD36, CD56) and glycophorin A
on RBCs.
4
CD 8 Purity
CD8 T cells are negatively selected from PBLs by
using antibodies directed against cell surface
antigens on human blood cells (CD4, CD14, CD16,
CD19, CD20, CD36, CD56, CD123, TCR gamma/sigma,
glycophorin A) and dextran.
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T Cell Purity
Black Isotype Blue CD3 Positive Cells
T Cells are negatively selected by using
antibodies targeting cell surface antigens on
human hematopoietic cells (CD16, CD19, CD36,
CD56) and glycophorin A on RBCs.
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Phenotype of T cell depleted Monocytes
SSC
CD11b
CD3 FITC
FSC
CD16 FITC
CD4 APC
CD14 PE
CD8 PE
HLA-DR FITC
CD3 FITC