hemacare products viability and functional data
Post on 18-Aug-2015
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Human-derived Primary Cells
2
Peripheral Blood
•Leukopaks
•Platelets
•Whole Blood
•Plasma
•Serum
•Mononuclear Cells
•CD3 Pan T Cells
•CD4 Helper T Cells
•CD8 Cytotoxic T Cells
•CD14 Monocytes
•CD19 B Cells
•CD4/CD25 Regulatory
T Cells
•CD56 NK Cells
•Dendritic Cells
•Macrophages
•Neutrophils
Mobilized Peripheral Blood
•Mononuclear Cells
•CD14 Monocytes
•CD34 Stem Cells
•CD34 Depleted
Mononuclear Cells
•CD133 Stem Cells
Bone Marrow
•Unprocessed Bone Marrow
•Mononuclear Cells
•Mesenchymal Stem Cells
•CD34 Stem Cells
•CD34 Depleted
Mononuclear Cells
•CD133 Stem Cells
Cord Blood
•Unprocessed Cord Blood
•Mononuclear Cells
•CD3 Pan T Cells
•CD4 Helper T Cells
•CD8 Cytotoxic T Cells
•CD14 Monocytes
•CD19 B Cells
•CD34 Stem Cells
•CD133 Stem Cells
•CD56 NK Cells
•Neutrophils
•Mesenchymal Stem Cells
•Plasma
•Dendritic Cells (CD34
derived)
High-quality Leukapheresis Raw
Material Collected at HemaCare Count (x106) ± SD
Total Nucleated Cells 14,400 550
Mononuclear Cells 12,100 600
4
n= 1,202 leukapheresis products
Peripheral Blood Mononuclear Cells (PBMCs)
6
Sources for Mononuclear cells
• Peripheral blood
• Mobilized peripheral blood
• Bone Marrow
• Cord blood
n = 20
Cryopreserved PBMC Functional Assay:
CMV Culture
7
INF-γ and TNF-α intracellular staining on day 6 PBMCs stimulated with CMV peptide
T-Cells
8
Sources for T-Cells
• Peripheral blood
• Mobilized peripheral blood
• Bone Marrow
• Cord blood
n = 6
National Institute of Allergy and Infectious
Diseases (NIAID)
CSFE Labeled CD3+ Pan T-cells
Cultured with Dendritic Cells:
Functional Assay
9
CFSE labeled T cells stimulated
by monocyte derived DC after 5
days undergo multiple divisions.
Ki67 and CD25 expression on T
cells stimulated by monocyte
derived DC after 5 days.
Natural Killer (NK) Cells
10
Sources for NK Cells
• Peripheral blood
• Cord blood
n = 7
0
20
40
60
80
100
Pre-Cryo Post-Thaw
% Purity
96.3% 93.0%
Monocytes
13
Sources for Monocytes
• Peripheral blood
• Mobilized peripheral blood
• Cord blood
n = 6
Macrophages (M1 and M2, monocyte derived)
14
n = 5
Source: National Cancer Institute (NCI); Creator: Sriram Subramaniam
Macrophages M1
15
Type 1-Macrophages (M1) were derived from purified Monocytes. CD14 monocytes were positively selected via
immunomagnetic beads and were cultured in the presence of GM-CSF for 10 days.
Macrophages M2
16
Type 2-Macrophages (M2) were derived from purified Monocytes. CD14 monocytes were positively selected via
immunomagnetic beads and were cultured in the presence of M-CSF for 10 days.
Dendritic Cells
17
Sources for Dendritic Cells
• Peripheral blood
• Cord blood
Quality data on next slide
Source: National Cancer Institute (NCI); Creator: Sriram Subramaniam
Dendritic Cells – Certificate of Analysis
18
Dendritic Cells (DC) were derived from purified CD14+ Monocytes. CD14+ cells were positively selected via
immunomagnetic beads and cultured in the presence of GM-CSF and IL-4 for 7 days
Neutrophils
19
Sources for Neutrophils
• Peripheral blood
• Cord blood
Source: National Institute of Allergy and Infectious Diseases (NIAID)
Stem/Progenitor Cell Products
20
Sources for Stem/Progenitor Cells
• Mobilized peripheral blood
• Bone Marrow
• Cord blood
n = 5
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