PMID- 11452164
OWN - NLM
STAT- MEDLINE
DCOM- 20011011
LR  - 20191210
IS  - 0041-1132 (Print)
IS  - 0041-1132 (Linking)
VI  - 41
IP  - 7
DP  - 2001 Jul
TI  - Ex vivo evaluation of PBMNCs collected with a new cell separator.
PG  - 940-9
AB  - BACKGROUND: This study reports on an evaluation of the ability of a cell 
      separator (Amicus, Baxter Healthcare) and the integral MNC computer software 
      program to collect a variety of MNC subsets. The collection efficiency (CE) of 
      the Amicus for these MNC subsets was compared to that of another cell separator 
      (CS-3000 Plus, Baxter). The collected MNCs were also assayed ex vivo to determine 
      if these cells remained functional. STUDY DESIGN AND METHODS: Healthy volunteer 
      blood donors were recruited to provide PBMNCs for the isolation of CD3+, CD4+, 
      CD8+, CD19+, NK, and gammadelta+ cells and monocytes. Cells were collected with 
      an Amicus (test arm; n = 16) or a CS-3000 Plus (control arm; n = 11) cell 
      separator. Cells were counted on a flow cytometer and CEs were calculated. For 
      functional studies, the Amicus-collected MNC data were compared to CS-3000 Plus 
      historical data. Functional studies performed included surface antigen expression 
      assays (CD8+), proliferation assays (CD4+ and CD8+ cells), NK cytotoxicity assays 
      for K562 and HUVE cells, and E-selectin induction on endothelial cells through 
      NK+ contact dependency. Dendritic cells (DCs) were generated from CD34+ cells 
      collected on the Amicus, positively selected by the use of antibody-bound, 
      magnetic bead technology, and then cultured ex vivo with a combination of growth 
      factors to generate the DCs. RESULTS: CEs were higher on the Amicus than on the 
      CS-3000 Plus for CD3+ (68 vs. 54%), CD4+ (70 vs. 56%), CD8+ (68 vs. 52%), and 
      CD19+ (60 vs. 48%) cells (p<0.05). For the two separators, CEs were equivalent 
      for monocytes, NK+, and gammadelta+ cells. The Amicus separator collected 
      significantly fewer platelets than did the CS-3000 Plus (p<0.00001). CD4+, CD8+, 
      and NK cells proliferated normally. NK cells appropriately stimulated E-selectin 
      expression on endothelial cells. Culture-generated DCs obtained by using 
      Amicus-collected CD34+ cells expressed appropriate cell surface markers. 
      CONCLUSION: The Amicus separator is acceptable for the collection of PBMNC 
      subsets. The device collects CD3+, CD4+, CD8+, and CD19+ T- and B-cell subsets 
      with greater efficiency and collects MNCs with significantly fewer contaminating 
      platelets than does the CS-3000 Plus. Cells collected on the Amicus are suitable 
      for use in a variety of research and clinical immunobiologic studies.
FAU - Snyder, E L
AU  - Snyder EL
AD  - Department of Laboratory Medicine, Yale University School of Medicine, New Haven, 
      Connecticut 06504, USA. edward.snyder@yale.edu
FAU - O'Donnell, L
AU  - O'Donnell L
FAU - Dengler, T J
AU  - Dengler TJ
FAU - Pomper, G J
AU  - Pomper GJ
FAU - Velleca, M A
AU  - Velleca MA
FAU - Dincecco, D M
AU  - Dincecco DM
FAU - Baril, L L
AU  - Baril LL
FAU - Min, K
AU  - Min K
FAU - Gudino, M D
AU  - Gudino MD
FAU - Bender, J R
AU  - Bender JR
LA  - eng
GR  - K08 AI01493/AI/NIAID NIH HHS/United States
GR  - R01 HL-43331/HL/NHLBI NIH HHS/United States
GR  - R01 HL-62223/HL/NHLBI NIH HHS/United States
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Transfusion
JT  - Transfusion
JID - 0417360
RN  - 0 (E-Selectin)
SB  - IM
MH  - Blood Donors
MH  - CD4-Positive T-Lymphocytes/cytology
MH  - CD8-Positive T-Lymphocytes/cytology
MH  - Cell Division/physiology
MH  - Cell Separation/*instrumentation
MH  - Cytotoxicity Tests, Immunologic
MH  - E-Selectin/biosynthesis
MH  - Endothelium, Vascular/cytology
MH  - Hematopoietic Stem Cell Transplantation
MH  - Humans
MH  - Infant, Newborn
MH  - Killer Cells, Natural/physiology
MH  - Leukapheresis
MH  - Leukocytes, Mononuclear/*cytology/physiology
MH  - Time Factors
MH  - Umbilical Veins
EDAT- 2001/07/14 10:00
MHDA- 2001/10/12 10:01
CRDT- 2001/07/14 10:00
PHST- 2001/07/14 10:00 [pubmed]
PHST- 2001/10/12 10:01 [medline]
PHST- 2001/07/14 10:00 [entrez]
AID - 10.1046/j.1537-2995.2001.41070940.x [doi]
PST - ppublish
SO  - Transfusion. 2001 Jul;41(7):940-9. doi: 10.1046/j.1537-2995.2001.41070940.x.