Abstract
The basic principles of cryobiology are discussed and applications of these principles by numerous workers in attempts to preserve marrow cells, leucocytes and platelets at low temperatures are reviewed. It is concluded that: (1) Lymphocytes from animals and man can be stored for long periods of time at low temperatures when cooled slowly in 10-15% DMSO or glycerol and thawed rapidly. Recovery figures are high and function is intact and unaltered. DMSO is probably a better preservative than glycerol, and storage life at -196 Degree-Sign C is probably indefinite for all practical purposes. Other leucocytes can be stored with similar techniques but recoveries after freezing and thawing are probably lower than with lymphocytes. (2) Bone-marrow cells of several animals including mouse, rabbit and dog can be preserved at -196 Degree-Sign C, probably indefinitely, and similar procedures to those used for lymphocytes give the best results. The comprehensive studies of Lewis and Trobaugh indicate that under carefully controlled conditions 95% of the stem cells in mouse marrow are viable after freezing and thawing. Opinions are divided over the efficacy of the two preservatives, DMSO and glycerol, but in view of the well documented accounts of the lack of toxicity of glycerol
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Ashwood-Smith, M. J.
[1]
- Medical Research Council, Radiobiological Research Unit, Harwell, Berks. (United Kingdom)
Citation Formats
Ashwood-Smith, M. J.
Preservation of Bone-Marrow Cells, Leucocytes and Platelets at Low Temperatures. A Review.
IAEA: N. p.,
1969.
Web.
Ashwood-Smith, M. J.
Preservation of Bone-Marrow Cells, Leucocytes and Platelets at Low Temperatures. A Review.
IAEA.
Ashwood-Smith, M. J.
1969.
"Preservation of Bone-Marrow Cells, Leucocytes and Platelets at Low Temperatures. A Review."
IAEA.
@misc{etde_22192478,
title = {Preservation of Bone-Marrow Cells, Leucocytes and Platelets at Low Temperatures. A Review}
author = {Ashwood-Smith, M. J.}
abstractNote = {The basic principles of cryobiology are discussed and applications of these principles by numerous workers in attempts to preserve marrow cells, leucocytes and platelets at low temperatures are reviewed. It is concluded that: (1) Lymphocytes from animals and man can be stored for long periods of time at low temperatures when cooled slowly in 10-15% DMSO or glycerol and thawed rapidly. Recovery figures are high and function is intact and unaltered. DMSO is probably a better preservative than glycerol, and storage life at -196 Degree-Sign C is probably indefinite for all practical purposes. Other leucocytes can be stored with similar techniques but recoveries after freezing and thawing are probably lower than with lymphocytes. (2) Bone-marrow cells of several animals including mouse, rabbit and dog can be preserved at -196 Degree-Sign C, probably indefinitely, and similar procedures to those used for lymphocytes give the best results. The comprehensive studies of Lewis and Trobaugh indicate that under carefully controlled conditions 95% of the stem cells in mouse marrow are viable after freezing and thawing. Opinions are divided over the efficacy of the two preservatives, DMSO and glycerol, but in view of the well documented accounts of the lack of toxicity of glycerol it would seem advisable for the moment to use this agent with all human marrow samples. The usefulness of PVP as a preservative for marrow is still to be resolved. Human marrow after freezing and thawing probably behaves in a similar manner to mouse marrow both in vitro and in vivo. However, it would be wise to consider that there might be differences which could cause wrong assessments of freezing procedures. (3) Platelet preservation, clinically perhaps the most useful procedure discussed in this review, is still to a large extent in the experimental stage. Much work has been done but even the best methods available permit relatively low recoveries of viable platelets. Preservation of human platelets in 12% glycerol associated with slow freezing and fast thawing and followed by very careful deglycerolization procedures yields a product with 23% of the original activity. This is clinically acceptable perhaps, but biologically it leaves much to be desired. Studies with rat platelets and combinations of various protective agents such as dextrose and DMSO and dextrose and dimethyl acetamide are encouraging, but toxicity problems may prove difficult in human studies. (4) The evidence from many different bacterial and animal cell studies indicates that -79 Degree-Sign C is not low enough for long-term preservation. Temperatures below -130 Degree-Sign C should be used whenever possible and precautions taken to exclude radiation from natural and cosmic sources when preservation in terms of years is considered. (author)}
place = {IAEA}
year = {1969}
month = {Jul}
}
title = {Preservation of Bone-Marrow Cells, Leucocytes and Platelets at Low Temperatures. A Review}
author = {Ashwood-Smith, M. J.}
abstractNote = {The basic principles of cryobiology are discussed and applications of these principles by numerous workers in attempts to preserve marrow cells, leucocytes and platelets at low temperatures are reviewed. It is concluded that: (1) Lymphocytes from animals and man can be stored for long periods of time at low temperatures when cooled slowly in 10-15% DMSO or glycerol and thawed rapidly. Recovery figures are high and function is intact and unaltered. DMSO is probably a better preservative than glycerol, and storage life at -196 Degree-Sign C is probably indefinite for all practical purposes. Other leucocytes can be stored with similar techniques but recoveries after freezing and thawing are probably lower than with lymphocytes. (2) Bone-marrow cells of several animals including mouse, rabbit and dog can be preserved at -196 Degree-Sign C, probably indefinitely, and similar procedures to those used for lymphocytes give the best results. The comprehensive studies of Lewis and Trobaugh indicate that under carefully controlled conditions 95% of the stem cells in mouse marrow are viable after freezing and thawing. Opinions are divided over the efficacy of the two preservatives, DMSO and glycerol, but in view of the well documented accounts of the lack of toxicity of glycerol it would seem advisable for the moment to use this agent with all human marrow samples. The usefulness of PVP as a preservative for marrow is still to be resolved. Human marrow after freezing and thawing probably behaves in a similar manner to mouse marrow both in vitro and in vivo. However, it would be wise to consider that there might be differences which could cause wrong assessments of freezing procedures. (3) Platelet preservation, clinically perhaps the most useful procedure discussed in this review, is still to a large extent in the experimental stage. Much work has been done but even the best methods available permit relatively low recoveries of viable platelets. Preservation of human platelets in 12% glycerol associated with slow freezing and fast thawing and followed by very careful deglycerolization procedures yields a product with 23% of the original activity. This is clinically acceptable perhaps, but biologically it leaves much to be desired. Studies with rat platelets and combinations of various protective agents such as dextrose and DMSO and dextrose and dimethyl acetamide are encouraging, but toxicity problems may prove difficult in human studies. (4) The evidence from many different bacterial and animal cell studies indicates that -79 Degree-Sign C is not low enough for long-term preservation. Temperatures below -130 Degree-Sign C should be used whenever possible and precautions taken to exclude radiation from natural and cosmic sources when preservation in terms of years is considered. (author)}
place = {IAEA}
year = {1969}
month = {Jul}
}