Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/7500
Citations | ||
Scopus | Web of ScienceĀ® | Altmetric |
---|---|---|
?
|
?
|
Type: | Journal article |
Title: | Removal of erythroid cells from umbilical cord blood mononuclear cell preparations using magnetic beads and a monoclonal antibody against glycophorin A |
Author: | Elliott, S. Macardle, P. Zola, H. |
Citation: | Journal of Immunological Methods, 1998; 217(1-2):121-130 |
Publisher: | Elsevier BV |
Issue Date: | 1998 |
ISSN: | 0022-1759 1872-7905 |
Abstract: | Umbilical cord blood mononuclear cells isolated by density centrifugation are contaminated by erythrocytes and nucleated erythroid precursors which may exceed 50% of the total cell population, and thus interfere with phenotypic, functional and mRNA analyses. Lysis with hypotonic ammonium chloride can overcome this problem, but interferes with lysosomal function and should be avoided when cell preparations are intended for functional studies. The aim of this study was to develop a technique for removing erythroid cells from cord blood mononuclear cell preparations that would be as effective as ammonium chloride lysis but would not affect cellular function. This paper describes a method using 10F7, a mouse monoclonal antibody against human glycophorin A, and magnetic beads coated with anti-mouse immunoglobulin. The population of cord blood mononuclear cells recovered using this technique was of high purity, good yield and viability, and the cells responded appropriately to stimulation in vitro. To maximise cost-effectiveness, purification with magnetic beads could be performed after two density separations to reduce the quantity of beads required. |
Keywords: | Erythrocytes Leukocytes, Mononuclear Fetal Blood Animals Humans Mice Ammonium Chloride Antibodies, Monoclonal Hypotonic Solutions Osmotic Fragility Centrifugation, Density Gradient Cell Separation Immunomagnetic Separation Infant, Newborn Evaluation Studies as Topic Glycophorins |
DOI: | 10.1016/S0022-1759(98)00111-2 |
Published version: | http://dx.doi.org/10.1016/s0022-1759(98)00111-2 |
Appears in Collections: | Aurora harvest 5 Paediatrics publications |
Files in This Item:
There are no files associated with this item.
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.