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Freeze-Drying of Mononuclear Cells Derived from Umbilical Cord Blood Followed by Colony Formation

BACKGROUND: We recently showed that freeze-dried cells stored for 3 years at room temperature can direct embryonic development following cloning. However, viability, as evaluated by membrane integrity of the cells after freeze-drying, was very low; and it was mainly the DNA integrity that was preser...

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Autores principales: Natan, Dity, Nagler, Arnon, Arav, Amir
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2667668/
https://www.ncbi.nlm.nih.gov/pubmed/19381290
http://dx.doi.org/10.1371/journal.pone.0005240
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author Natan, Dity
Nagler, Arnon
Arav, Amir
author_facet Natan, Dity
Nagler, Arnon
Arav, Amir
author_sort Natan, Dity
collection PubMed
description BACKGROUND: We recently showed that freeze-dried cells stored for 3 years at room temperature can direct embryonic development following cloning. However, viability, as evaluated by membrane integrity of the cells after freeze-drying, was very low; and it was mainly the DNA integrity that was preserved. In the present study, we improved the cells' viability and functionality after freeze-drying. METHODOLOGY/PRINCIPAL FINDINGS: We optimized the conditions of directional freezing, i.e. interface velocity and cell concentration, and we added the antioxidant EGCG to the freezing solution. The study was performed on mononuclear cells (MNCs) derived from human umbilical cord blood. After freeze-drying, we tested the viability, number of CD34(+)-presenting cells and ability of the rehydrated hematopoietic stem cells to differentiate into different blood cells in culture. The viability of the MNCs after freeze-drying and rehydration with pure water was 88%–91%. The total number of CD34(+)-presenting cells and the number of colonies did not change significantly when evaluated before freezing, after freeze-thawing, and after freeze-drying (5.4×10(4)±4.7, 3.49×10(4)±6 and 6.31×10(4)±12.27 cells, respectively, and 31±25.15, 47±45.8 and 23.44±13.3 colonies, respectively). CONCLUSIONS: This is the first report of nucleated cells which have been dried and then rehydrated with double-distilled water remaining viable, and of hematopoietic stem cells retaining their ability to differentiate into different blood cells.
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spelling pubmed-26676682009-04-21 Freeze-Drying of Mononuclear Cells Derived from Umbilical Cord Blood Followed by Colony Formation Natan, Dity Nagler, Arnon Arav, Amir PLoS One Research Article BACKGROUND: We recently showed that freeze-dried cells stored for 3 years at room temperature can direct embryonic development following cloning. However, viability, as evaluated by membrane integrity of the cells after freeze-drying, was very low; and it was mainly the DNA integrity that was preserved. In the present study, we improved the cells' viability and functionality after freeze-drying. METHODOLOGY/PRINCIPAL FINDINGS: We optimized the conditions of directional freezing, i.e. interface velocity and cell concentration, and we added the antioxidant EGCG to the freezing solution. The study was performed on mononuclear cells (MNCs) derived from human umbilical cord blood. After freeze-drying, we tested the viability, number of CD34(+)-presenting cells and ability of the rehydrated hematopoietic stem cells to differentiate into different blood cells in culture. The viability of the MNCs after freeze-drying and rehydration with pure water was 88%–91%. The total number of CD34(+)-presenting cells and the number of colonies did not change significantly when evaluated before freezing, after freeze-thawing, and after freeze-drying (5.4×10(4)±4.7, 3.49×10(4)±6 and 6.31×10(4)±12.27 cells, respectively, and 31±25.15, 47±45.8 and 23.44±13.3 colonies, respectively). CONCLUSIONS: This is the first report of nucleated cells which have been dried and then rehydrated with double-distilled water remaining viable, and of hematopoietic stem cells retaining their ability to differentiate into different blood cells. Public Library of Science 2009-04-21 /pmc/articles/PMC2667668/ /pubmed/19381290 http://dx.doi.org/10.1371/journal.pone.0005240 Text en Natan et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Natan, Dity
Nagler, Arnon
Arav, Amir
Freeze-Drying of Mononuclear Cells Derived from Umbilical Cord Blood Followed by Colony Formation
title Freeze-Drying of Mononuclear Cells Derived from Umbilical Cord Blood Followed by Colony Formation
title_full Freeze-Drying of Mononuclear Cells Derived from Umbilical Cord Blood Followed by Colony Formation
title_fullStr Freeze-Drying of Mononuclear Cells Derived from Umbilical Cord Blood Followed by Colony Formation
title_full_unstemmed Freeze-Drying of Mononuclear Cells Derived from Umbilical Cord Blood Followed by Colony Formation
title_short Freeze-Drying of Mononuclear Cells Derived from Umbilical Cord Blood Followed by Colony Formation
title_sort freeze-drying of mononuclear cells derived from umbilical cord blood followed by colony formation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2667668/
https://www.ncbi.nlm.nih.gov/pubmed/19381290
http://dx.doi.org/10.1371/journal.pone.0005240
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