Comparison of purity and enrichment of CD34+ cells from bone marrow, umbilical cord and peripheral blood (primed for apheresis) using five separation systems.

2.50
Hdl Handle:
http://hdl.handle.net/10541/97963
Title:
Comparison of purity and enrichment of CD34+ cells from bone marrow, umbilical cord and peripheral blood (primed for apheresis) using five separation systems.
Authors:
De Wynter, Erika A; Coutinho, Lucia H; Pei, X; Marsh, J C; Hows, J; Luft, T; Testa, Nydia G
Abstract:
Interest in the isolation and characterization of primitive hemopoietic cells in both the clinical and research fields has rapidly increased. In parallel, different purification systems have been developed to isolate these cells. We have compared five different methods for separation of CD34+ cells from human umbilical cord blood, normal bone marrow and apheresis harvests and analyzed purity, recovery, yield and enrichment of colony forming cells (CFC) for each individual system. Our results indicate that the most reliable methods of purification for all samples were fluorescence activated cell sorting (FACS) and magnetic activated cell sorting (MACS) which consistently yielded high purities (> 70%) and enrichment of CFC. In this respect the enrichment of CFC from the MACS was superior to all the other systems including FACS. Similar results (> 70%) for purity were obtained using avidin affinity columns and a biotinylated antibody but neither yield nor CFC enrichment approached the values achieved with MACS. On average CFC enrichment using these affinity columns was greater than that observed for FACS while the purity was comparable. Both CELLector flasks and immunomagnetic beads coated with CD34 antibodies were less effective in our hands in separating purified populations of progenitor cells. Both purity and CFC enrichment of CD34+ cells using these methods were at least 50% lower than obtained with either FACS, MACS or affinity columns.
Affiliation:
Cancer Research Campaign Department of Experimental Haematology, Paterson Institute for Cancer Research, Manchester, United Kingdom.
Citation:
Comparison of purity and enrichment of CD34+ cells from bone marrow, umbilical cord and peripheral blood (primed for apheresis) using five separation systems. 1995, 13 (5):524-32 Stem Cells
Journal:
Stem Cells
Issue Date:
Sep-1995
URI:
http://hdl.handle.net/10541/97963
DOI:
10.1002/stem.5530130510
PubMed ID:
8528102
Type:
Article
Language:
en
ISSN:
1066-5099
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorDe Wynter, Erika Aen
dc.contributor.authorCoutinho, Lucia Hen
dc.contributor.authorPei, Xen
dc.contributor.authorMarsh, J Cen
dc.contributor.authorHows, Jen
dc.contributor.authorLuft, Ten
dc.contributor.authorTesta, Nydia Gen
dc.date.accessioned2010-05-05T10:42:43Z-
dc.date.available2010-05-05T10:42:43Z-
dc.date.issued1995-09-
dc.identifier.citationComparison of purity and enrichment of CD34+ cells from bone marrow, umbilical cord and peripheral blood (primed for apheresis) using five separation systems. 1995, 13 (5):524-32 Stem Cellsen
dc.identifier.issn1066-5099-
dc.identifier.pmid8528102-
dc.identifier.doi10.1002/stem.5530130510-
dc.identifier.urihttp://hdl.handle.net/10541/97963-
dc.description.abstractInterest in the isolation and characterization of primitive hemopoietic cells in both the clinical and research fields has rapidly increased. In parallel, different purification systems have been developed to isolate these cells. We have compared five different methods for separation of CD34+ cells from human umbilical cord blood, normal bone marrow and apheresis harvests and analyzed purity, recovery, yield and enrichment of colony forming cells (CFC) for each individual system. Our results indicate that the most reliable methods of purification for all samples were fluorescence activated cell sorting (FACS) and magnetic activated cell sorting (MACS) which consistently yielded high purities (> 70%) and enrichment of CFC. In this respect the enrichment of CFC from the MACS was superior to all the other systems including FACS. Similar results (> 70%) for purity were obtained using avidin affinity columns and a biotinylated antibody but neither yield nor CFC enrichment approached the values achieved with MACS. On average CFC enrichment using these affinity columns was greater than that observed for FACS while the purity was comparable. Both CELLector flasks and immunomagnetic beads coated with CD34 antibodies were less effective in our hands in separating purified populations of progenitor cells. Both purity and CFC enrichment of CD34+ cells using these methods were at least 50% lower than obtained with either FACS, MACS or affinity columns.en
dc.language.isoenen
dc.subjectFoetal Blooden
dc.subjectHaematopoietic Stem Cellsen
dc.subject.meshAntigens, CD-
dc.subject.meshAntigens, CD34-
dc.subject.meshBlood Cells-
dc.subject.meshBlood Component Removal-
dc.subject.meshBone Marrow Cells-
dc.subject.meshCell Separation-
dc.subject.meshCells, Cultured-
dc.subject.meshCulture Techniques-
dc.subject.meshFemale-
dc.subject.meshFetal Blood-
dc.subject.meshFlow Cytometry-
dc.subject.meshHematopoietic Stem Cells-
dc.subject.meshHumans-
dc.subject.meshImmunomagnetic Separation-
dc.subject.meshInfant, Newborn-
dc.subject.meshPregnancy-
dc.titleComparison of purity and enrichment of CD34+ cells from bone marrow, umbilical cord and peripheral blood (primed for apheresis) using five separation systems.en
dc.typeArticleen
dc.contributor.departmentCancer Research Campaign Department of Experimental Haematology, Paterson Institute for Cancer Research, Manchester, United Kingdom.en
dc.identifier.journalStem Cellsen

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