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A comparison of haematopoietic progenitor cells enumeration using sysmex haematology analyzer and standard flow cytometry

The CD34+ antigen is present on immature haematopoietic progenitor cells and all haematopoietic colony-forming cells in bone marrow and blood. 'The administration of recombinant human granulocyte colony-stimulating factor (rhG-CSF) has emerged as an efficient way to accomplish mobilization per...

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Main Author: Zulkafli, Zefarina
Format: Thesis
Language:English
Published: 2010
Subjects:
QH573-671 Cytology
Online Access:http://eprints.usm.my/58932/1/DR%20ZEFARINA%20BINTI%20ZULKIFLI%20-%20e24.pdf
http://eprints.usm.my/58932/
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institution Universiti Sains Malaysia
building Hamzah Sendut Library
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continent Asia
country Malaysia
content_provider Universiti Sains Malaysia
content_source USM Institutional Repository
url_provider http://eprints.usm.my/
language English
topic QH573-671 Cytology
spellingShingle QH573-671 Cytology
Zulkafli, Zefarina
A comparison of haematopoietic progenitor cells enumeration using sysmex haematology analyzer and standard flow cytometry
description The CD34+ antigen is present on immature haematopoietic progenitor cells and all haematopoietic colony-forming cells in bone marrow and blood. 'The administration of recombinant human granulocyte colony-stimulating factor (rhG-CSF) has emerged as an efficient way to accomplish mobilization peripheral blood stem cells (PBSCs). The decision to harvest PBSC is based on the enumeration of CD 34 cells by flow cytometry in which require daily CD34+ monitoring after given chemotherapy-based mobilization regimens. Currently, CD34+ enumeration will be initiated when the white blood cell (WBC) count is more than 1 X 10J /pL. Flow cytometry assays have been used as a standard method for CD34+ enumeration in leukapheresis products before peripheral stem cell collection. However, flow cytometry techniques are time consuming, complex and expensive. Recently, technology has become available on a routine haematology analyzer that enables the detection of hematopoietic progenitor cells (HPC). Detection and enumeration of HPC by Sysmex haematology analyzer could provide a standard, cost effective and rapid alternative for predicting the yield of stem cells. The aims of this study were 1) To compare Haematopoietic Progenitor cells (HPC) enumeration by using Sysmex haematology analyzer and CD34+ count by standard flow cytometry in cord blood. ; 2) To correlate the number of CD34+ cells by standard flow cytometry with Haematopoietic Progenitor cells (HPC), white blood cells (WBC), immature platelet fraction (IPF), Reticulocyte (retie) and immature reticulocyte fraction (IRF) acquired by Sysmex haematology analyzer in patient on Granulocyte growth stimulating factor ( G-CSF) and 3) to predict the level of CD34+ cells by using the haematological parameters derived from Sysmex haematology analyzer. A cross sectional study was done in Stem Cell laboratory at HUSM from November 2008 to June 2009. 3mls of EDTA anticoagulated blood from cord blood of 95 newborns were collected and 68 samples of 3mls EDTA anticoagulated blood were collected from 19 adult patients with haematological malignancies from HUSM who had received chemotherapybased mobilization regimens including G-CSF. Progenitor cell quantification was performed measuring HPC counts as well as other haematological parameters, WBC, IPF, RETIC and IRF counts provided by the Sysmex XE-2100 haematology analyzer and CD34+ counts obtained in parallel by flow cytometry. Data were analyzed using the Statistical Packages for Social Sciences (SPSS) software version 12.0. Analysis of the umbilical cord blood showed that mean(SD) for HPC count was 32.6 (34.3) X 106/ L and mean(SD) for CD34+ count was 36.8 (27.2) X 106/ L. There was no significant difference between mean HPC count and CD34+ count. Blood samples from adult patient with haematological malignancies, were analysed by Spearman correlation, showed a highly significant correlation between CD34+ and HPC count (r = 0.703 , pO.OOl), WBC ( r = 0.482 , pO.OOl), Retie ( r = 0.498 , pO.OOl) and IPF (r = 0.453 , pO.OOl) but not with IRF ( r = -0.073 , p = 0.552). The other objective was to determined the optimal cut-off point for the haematological parameters that predicts CD34+ count of > 20 X 106/ L. This is because in current practice the harvest will be initiated when CD34+ count is > 20 X 106/ L. The result showed that HPC, IPF, Retie, WBC and IRF threshold were > 21.5 X 106/L, > 26.1%, > 1% , > 23.1 X 109/L and > 8.9% respectively wirh sensitivity and specificity were (77% and 64%, 77% and 50%, 73% and 64% , 73% and 50% and 52% respectively). In this study we found that, there is a significant correlation between the number of CD34+ cells measured using flow cytometry with HPC, RETIC, WBC, IPF measured by Sysmex haematology analyzer but not with IRF. This study revealed that HPC, WBC, RETIC and IPF counts can be applied independently into clinical protocol to evaluate the timing of leukapheresis for starting PBSC collection.
format Thesis
author Zulkafli, Zefarina
author_facet Zulkafli, Zefarina
author_sort Zulkafli, Zefarina
title A comparison of haematopoietic progenitor cells enumeration using sysmex haematology analyzer and standard flow cytometry
title_short A comparison of haematopoietic progenitor cells enumeration using sysmex haematology analyzer and standard flow cytometry
title_full A comparison of haematopoietic progenitor cells enumeration using sysmex haematology analyzer and standard flow cytometry
title_fullStr A comparison of haematopoietic progenitor cells enumeration using sysmex haematology analyzer and standard flow cytometry
title_full_unstemmed A comparison of haematopoietic progenitor cells enumeration using sysmex haematology analyzer and standard flow cytometry
title_sort comparison of haematopoietic progenitor cells enumeration using sysmex haematology analyzer and standard flow cytometry
publishDate 2010
url http://eprints.usm.my/58932/1/DR%20ZEFARINA%20BINTI%20ZULKIFLI%20-%20e24.pdf
http://eprints.usm.my/58932/
_version_ 1775623409637523456
spelling my.usm.eprints.58932 http://eprints.usm.my/58932/ A comparison of haematopoietic progenitor cells enumeration using sysmex haematology analyzer and standard flow cytometry Zulkafli, Zefarina QH573-671 Cytology The CD34+ antigen is present on immature haematopoietic progenitor cells and all haematopoietic colony-forming cells in bone marrow and blood. 'The administration of recombinant human granulocyte colony-stimulating factor (rhG-CSF) has emerged as an efficient way to accomplish mobilization peripheral blood stem cells (PBSCs). The decision to harvest PBSC is based on the enumeration of CD 34 cells by flow cytometry in which require daily CD34+ monitoring after given chemotherapy-based mobilization regimens. Currently, CD34+ enumeration will be initiated when the white blood cell (WBC) count is more than 1 X 10J /pL. Flow cytometry assays have been used as a standard method for CD34+ enumeration in leukapheresis products before peripheral stem cell collection. However, flow cytometry techniques are time consuming, complex and expensive. Recently, technology has become available on a routine haematology analyzer that enables the detection of hematopoietic progenitor cells (HPC). Detection and enumeration of HPC by Sysmex haematology analyzer could provide a standard, cost effective and rapid alternative for predicting the yield of stem cells. The aims of this study were 1) To compare Haematopoietic Progenitor cells (HPC) enumeration by using Sysmex haematology analyzer and CD34+ count by standard flow cytometry in cord blood. ; 2) To correlate the number of CD34+ cells by standard flow cytometry with Haematopoietic Progenitor cells (HPC), white blood cells (WBC), immature platelet fraction (IPF), Reticulocyte (retie) and immature reticulocyte fraction (IRF) acquired by Sysmex haematology analyzer in patient on Granulocyte growth stimulating factor ( G-CSF) and 3) to predict the level of CD34+ cells by using the haematological parameters derived from Sysmex haematology analyzer. A cross sectional study was done in Stem Cell laboratory at HUSM from November 2008 to June 2009. 3mls of EDTA anticoagulated blood from cord blood of 95 newborns were collected and 68 samples of 3mls EDTA anticoagulated blood were collected from 19 adult patients with haematological malignancies from HUSM who had received chemotherapybased mobilization regimens including G-CSF. Progenitor cell quantification was performed measuring HPC counts as well as other haematological parameters, WBC, IPF, RETIC and IRF counts provided by the Sysmex XE-2100 haematology analyzer and CD34+ counts obtained in parallel by flow cytometry. Data were analyzed using the Statistical Packages for Social Sciences (SPSS) software version 12.0. Analysis of the umbilical cord blood showed that mean(SD) for HPC count was 32.6 (34.3) X 106/ L and mean(SD) for CD34+ count was 36.8 (27.2) X 106/ L. There was no significant difference between mean HPC count and CD34+ count. Blood samples from adult patient with haematological malignancies, were analysed by Spearman correlation, showed a highly significant correlation between CD34+ and HPC count (r = 0.703 , pO.OOl), WBC ( r = 0.482 , pO.OOl), Retie ( r = 0.498 , pO.OOl) and IPF (r = 0.453 , pO.OOl) but not with IRF ( r = -0.073 , p = 0.552). The other objective was to determined the optimal cut-off point for the haematological parameters that predicts CD34+ count of > 20 X 106/ L. This is because in current practice the harvest will be initiated when CD34+ count is > 20 X 106/ L. The result showed that HPC, IPF, Retie, WBC and IRF threshold were > 21.5 X 106/L, > 26.1%, > 1% , > 23.1 X 109/L and > 8.9% respectively wirh sensitivity and specificity were (77% and 64%, 77% and 50%, 73% and 64% , 73% and 50% and 52% respectively). In this study we found that, there is a significant correlation between the number of CD34+ cells measured using flow cytometry with HPC, RETIC, WBC, IPF measured by Sysmex haematology analyzer but not with IRF. This study revealed that HPC, WBC, RETIC and IPF counts can be applied independently into clinical protocol to evaluate the timing of leukapheresis for starting PBSC collection. 2010 Thesis NonPeerReviewed application/pdf en http://eprints.usm.my/58932/1/DR%20ZEFARINA%20BINTI%20ZULKIFLI%20-%20e24.pdf Zulkafli, Zefarina (2010) A comparison of haematopoietic progenitor cells enumeration using sysmex haematology analyzer and standard flow cytometry. Masters thesis, Universiti Sains Malaysia.
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