Optimizing the recovery of peripheral blood mononuclear cells trapped in leukoreduction filters - A comparison study

Abstract Introduction The isolation of captured peripheral blood mononuclear cells (PBMNCs) from leukoreduction filters (LRFs) can be of great importance in terms of bringing the lost cells back into use. Objective The aim of this study was to evaluate various methods based on their potential to recover the peripheral blood cells from LRFs with a focus on mononuclear cells (MNCs). Method For cell isolation from LRFs, three distinct methods (back-flushing, direct and vacuum pump) were compared through the calculation of the yield of isolated MNCs. The viability of extracted cells was determined by the flow cytometry technique. Moreover, the recovered MNCs were characterized regarding the presence of blood stem cell purification. The cell culture, microscopic observation, and immunophenotyping were employed to characterize the blood stem cells (hematopoietic, mesenchymal and progenitor endothelial stem cells). Results The yield of isolation obtained in the back-flushing, direct and vacuum pump methods were 17.7 ± 1.28, 17.3 ± 0.96 and 21.2 ± 0.90 percent, respectively. Although the highest potential for total blood cell recovery belonged to the vacuum pump method, the lowest cell viability (85.73 ± 4.84%) was observed in this method. However, the isolation process of the back-flushing and direct methods had less effect on cell viability. The characterization of the isolated MNCs displayed that the dominant positive phenotype was for CD34/CD45, indicating hematopoietic stem cells. In addition, the endothelial stem/progenitor cells were significantly detected as CD31/CD133 positive cells. Conclusion According to our results and considering the safety and efficiency potential of each of the applied methods, the back-flushing in comparison with the other methods can be considered a suitable procedure for MNC isolation from LRFs.

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Main Authors: Bashiri Dezfouli,Ali, Pourfathollah,Ali Akbar, Nikougoftar-Zarif,Mahin, Khosravi,Mohammad, Tajrishi,Mona, Ezzati,Nasim, Kashani Khatib,Zahra, Abbasi Sourki,Parvaneh, Valizadeh,Maryam
Format: Digital revista
Language:English
Published: Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular (ABHH) 2022
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S2531-13792022000200197
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spelling oai:scielo:S2531-137920220002001972022-06-08Optimizing the recovery of peripheral blood mononuclear cells trapped in leukoreduction filters - A comparison studyBashiri Dezfouli,AliPourfathollah,Ali AkbarNikougoftar-Zarif,MahinKhosravi,MohammadTajrishi,MonaEzzati,NasimKashani Khatib,ZahraAbbasi Sourki,ParvanehValizadeh,Maryam Leukoreduction filters Isolation methods Mononuclear cells Peripheral blood stem cells Abstract Introduction The isolation of captured peripheral blood mononuclear cells (PBMNCs) from leukoreduction filters (LRFs) can be of great importance in terms of bringing the lost cells back into use. Objective The aim of this study was to evaluate various methods based on their potential to recover the peripheral blood cells from LRFs with a focus on mononuclear cells (MNCs). Method For cell isolation from LRFs, three distinct methods (back-flushing, direct and vacuum pump) were compared through the calculation of the yield of isolated MNCs. The viability of extracted cells was determined by the flow cytometry technique. Moreover, the recovered MNCs were characterized regarding the presence of blood stem cell purification. The cell culture, microscopic observation, and immunophenotyping were employed to characterize the blood stem cells (hematopoietic, mesenchymal and progenitor endothelial stem cells). Results The yield of isolation obtained in the back-flushing, direct and vacuum pump methods were 17.7 ± 1.28, 17.3 ± 0.96 and 21.2 ± 0.90 percent, respectively. Although the highest potential for total blood cell recovery belonged to the vacuum pump method, the lowest cell viability (85.73 ± 4.84%) was observed in this method. However, the isolation process of the back-flushing and direct methods had less effect on cell viability. The characterization of the isolated MNCs displayed that the dominant positive phenotype was for CD34/CD45, indicating hematopoietic stem cells. In addition, the endothelial stem/progenitor cells were significantly detected as CD31/CD133 positive cells. Conclusion According to our results and considering the safety and efficiency potential of each of the applied methods, the back-flushing in comparison with the other methods can be considered a suitable procedure for MNC isolation from LRFs.info:eu-repo/semantics/openAccessAssociação Brasileira de Hematologia, Hemoterapia e Terapia Celular (ABHH)Hematology, Transfusion and Cell Therapy v.44 n.2 20222022-04-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S2531-13792022000200197en10.1016/j.htct.2020.09.155
institution SCIELO
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countrycode BR
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libraryname SciELO
language English
format Digital
author Bashiri Dezfouli,Ali
Pourfathollah,Ali Akbar
Nikougoftar-Zarif,Mahin
Khosravi,Mohammad
Tajrishi,Mona
Ezzati,Nasim
Kashani Khatib,Zahra
Abbasi Sourki,Parvaneh
Valizadeh,Maryam
spellingShingle Bashiri Dezfouli,Ali
Pourfathollah,Ali Akbar
Nikougoftar-Zarif,Mahin
Khosravi,Mohammad
Tajrishi,Mona
Ezzati,Nasim
Kashani Khatib,Zahra
Abbasi Sourki,Parvaneh
Valizadeh,Maryam
Optimizing the recovery of peripheral blood mononuclear cells trapped in leukoreduction filters - A comparison study
author_facet Bashiri Dezfouli,Ali
Pourfathollah,Ali Akbar
Nikougoftar-Zarif,Mahin
Khosravi,Mohammad
Tajrishi,Mona
Ezzati,Nasim
Kashani Khatib,Zahra
Abbasi Sourki,Parvaneh
Valizadeh,Maryam
author_sort Bashiri Dezfouli,Ali
title Optimizing the recovery of peripheral blood mononuclear cells trapped in leukoreduction filters - A comparison study
title_short Optimizing the recovery of peripheral blood mononuclear cells trapped in leukoreduction filters - A comparison study
title_full Optimizing the recovery of peripheral blood mononuclear cells trapped in leukoreduction filters - A comparison study
title_fullStr Optimizing the recovery of peripheral blood mononuclear cells trapped in leukoreduction filters - A comparison study
title_full_unstemmed Optimizing the recovery of peripheral blood mononuclear cells trapped in leukoreduction filters - A comparison study
title_sort optimizing the recovery of peripheral blood mononuclear cells trapped in leukoreduction filters - a comparison study
description Abstract Introduction The isolation of captured peripheral blood mononuclear cells (PBMNCs) from leukoreduction filters (LRFs) can be of great importance in terms of bringing the lost cells back into use. Objective The aim of this study was to evaluate various methods based on their potential to recover the peripheral blood cells from LRFs with a focus on mononuclear cells (MNCs). Method For cell isolation from LRFs, three distinct methods (back-flushing, direct and vacuum pump) were compared through the calculation of the yield of isolated MNCs. The viability of extracted cells was determined by the flow cytometry technique. Moreover, the recovered MNCs were characterized regarding the presence of blood stem cell purification. The cell culture, microscopic observation, and immunophenotyping were employed to characterize the blood stem cells (hematopoietic, mesenchymal and progenitor endothelial stem cells). Results The yield of isolation obtained in the back-flushing, direct and vacuum pump methods were 17.7 ± 1.28, 17.3 ± 0.96 and 21.2 ± 0.90 percent, respectively. Although the highest potential for total blood cell recovery belonged to the vacuum pump method, the lowest cell viability (85.73 ± 4.84%) was observed in this method. However, the isolation process of the back-flushing and direct methods had less effect on cell viability. The characterization of the isolated MNCs displayed that the dominant positive phenotype was for CD34/CD45, indicating hematopoietic stem cells. In addition, the endothelial stem/progenitor cells were significantly detected as CD31/CD133 positive cells. Conclusion According to our results and considering the safety and efficiency potential of each of the applied methods, the back-flushing in comparison with the other methods can be considered a suitable procedure for MNC isolation from LRFs.
publisher Associação Brasileira de Hematologia, Hemoterapia e Terapia Celular (ABHH)
publishDate 2022
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S2531-13792022000200197
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