PMID- 19421174
OWN - NLM
STAT- MEDLINE
DCOM- 20100323
LR  - 20171116
IS  - 1476-5365 (Electronic)
IS  - 0268-3369 (Linking)
VI  - 45
IP  - 1
DP  - 2010 Jan
TI  - Enrichment of cell subpopulations applying automated MACS technique: purity, 
      recovery and applicability for PCR-based chimerism analysis.
PG  - 181-9
LID - 10.1038/bmt.2009.89 [doi]
AB  - Enrichment of cell subpopulations is a prerequisite for lineage-specific 
      chimerism analysis (LCA), a frequent approach in follow-up after allo-SCT. An 
      efficient enrichment technique is Magnetic Cell Sorting (MACS) using the AutoMACS 
      separator. However, evaluation of purity, recovery and applicability for 
      PCR-based chimerism analysis of MACS-enriched subpopulations from post-transplant 
      peripheral blood, providing reduced cell numbers and/or unbalanced proportions of 
      subpopulations, is currently unavailable. We performed enrichment of CD3-, CD14-, 
      CD15-, CD19- and CD56-positive subpopulations using 'Whole Blood MicroBeads' and 
      AutoMACS separator in 137 prospectively collected peripheral blood samples from 
      15 paediatric patients after allo-CD3-/CD19-depleted SCT. Purity was assessed by 
      immune phenotyping. Recovery and applicability for chimerism analysis was 
      evaluated. Excellent purity >90% was achieved in CD14-, CD15-positive cells in 
      81%, 95% of the isolates and in 86% of CD3 and CD19 isolates, if ACC was >400 
      cells per mul. Median purity of CD56-positive isolates was 78.9%. Recovery >90% 
      was between 93 (CD56) and 37% (CD15). Conventional and real-time PCR-based 
      chimerism analysis was feasible in virtually all samples. Isolation of cell 
      subpopulations by automated cell enrichment in post-transplant peripheral blood 
      is feasible and fast providing excellent purity and recovery for routine 
      lineage-specific chimerism analysis.
FAU - Willasch, A
AU  - Willasch A
AD  - Department of Pediatric Hematology, Oncology and Hemostaseology, Goethe 
      University Frankfurt, Hospital for Children and Adolescents III, Frankfurt, 
      Germany. andre.willasch@kgu.de
FAU - Eing, S
AU  - Eing S
FAU - Weber, G
AU  - Weber G
FAU - Kuci, S
AU  - Kuci S
FAU - Schneider, G
AU  - Schneider G
FAU - Soerensen, J
AU  - Soerensen J
FAU - Jarisch, A
AU  - Jarisch A
FAU - Rettinger, E
AU  - Rettinger E
FAU - Koehl, U
AU  - Koehl U
FAU - Klingebiel, T
AU  - Klingebiel T
FAU - Kreyenberg, H
AU  - Kreyenberg H
FAU - Bader, P
AU  - Bader P
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20090504
PL  - England
TA  - Bone Marrow Transplant
JT  - Bone marrow transplantation
JID - 8702459
RN  - 0 (Antigens, CD19)
RN  - 0 (CD3 Complex)
RN  - 0 (CD56 Antigen)
RN  - 0 (Lewis X Antigen)
RN  - 0 (Lipopolysaccharide Receptors)
SB  - IM
MH  - Adolescent
MH  - Adult
MH  - Antigens, CD19/immunology
MH  - CD3 Complex/immunology
MH  - CD56 Antigen/immunology
MH  - *Cell Lineage
MH  - Cell Separation/*methods
MH  - Child
MH  - Child, Preschool
MH  - Female
MH  - Flow Cytometry/methods
MH  - Hematopoietic Stem Cell Transplantation/*methods
MH  - Humans
MH  - Lewis X Antigen/immunology
MH  - Lipopolysaccharide Receptors/immunology
MH  - Lymphocyte Subsets/*immunology
MH  - Magnetics
MH  - Male
MH  - Polymerase Chain Reaction
MH  - Polymorphism, Genetic
MH  - Postoperative Care
MH  - Tandem Repeat Sequences
MH  - Transplantation Chimera/*immunology
EDAT- 2009/05/08 09:00
MHDA- 2010/03/24 06:00
CRDT- 2009/05/08 09:00
PHST- 2009/05/08 09:00 [entrez]
PHST- 2009/05/08 09:00 [pubmed]
PHST- 2010/03/24 06:00 [medline]
AID - bmt200989 [pii]
AID - 10.1038/bmt.2009.89 [doi]
PST - ppublish
SO  - Bone Marrow Transplant. 2010 Jan;45(1):181-9. doi: 10.1038/bmt.2009.89. Epub 2009 
      May 4.