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...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2009-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC2667668?pdf=render |
| _version_ | 1811310340046061568 |
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| author | Dity Natan Arnon Nagler Amir Arav |
| author_facet | Dity Natan Arnon Nagler Amir Arav |
| author_sort | Dity Natan |
| collection | DOAJ |
| 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 x 10(4)+/-4.7, 3.49 x 10(4)+/-6 and 6.31 x 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. |
| first_indexed | 2024-04-13T09:57:19Z |
| format | Article |
| id | doaj.art-31102317ea6b4e98a5e624e096976f71 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-13T09:57:19Z |
| publishDate | 2009-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-31102317ea6b4e98a5e624e096976f712022-12-22T02:51:19ZengPublic Library of Science (PLoS)PLoS ONE1932-62032009-01-0144e524010.1371/journal.pone.0005240Freeze-drying of mononuclear cells derived from umbilical cord blood followed by colony formation.Dity NatanArnon NaglerAmir AravBACKGROUND: 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 x 10(4)+/-4.7, 3.49 x 10(4)+/-6 and 6.31 x 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.http://europepmc.org/articles/PMC2667668?pdf=render |
| spellingShingle | Dity Natan Arnon Nagler Amir Arav Freeze-drying of mononuclear cells derived from umbilical cord blood followed by colony formation. PLoS ONE |
| 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 |
| url | http://europepmc.org/articles/PMC2667668?pdf=render |
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