MAJOR QUALITY PARAMETERS OF LONG-TERM CRYOPRESERVED CORD BLOOD UNITS - A SINGLE CENTER EXPERIENCE
(Abstract release date: 05/19/16)
EHA Library. Bordalo F. 06/09/16; 135087; PB2187
Mr. Filipa Bordalo
Contributions
Contributions
Abstract
Abstract: PB2187
Type: Publication Only
Background
Cord Blood (CB) is a well-recognized source of stem cells for hematopoietic transplant. Its main advantage is to be stored for long-term with a ready availability for the intended recipient and no risk for the donor.
Aims
The aim of this study was to evaluate the quality of 15 cord blood units cryopreserved in our direct donation bank. for the intended recipient and no risk for the donor.
Methods
All these units were processed without red cells reduction, in a 24 hours period after collection, then were cryopreserved with a controlled-rate freezer and transferred to a liquid nitrogen container. We used cryogenic bags (Kapton/Teflon), highly resistant to very low temperatures. However, due to missing data we do not considered these CB units suitable for transplant. In this study, the CB were thawed and washed based on the New York Blood Center method in our ISO 7 cleanroom, inside of an air flow cabinet. We performed cellular counts, viability assays (by flow cytometry using 7-aminoactinomycin), sterility testing (bacterial and fungal cultures) and clonogenicity (colony-forming units-granulocyte/macrophage - CFU-GM - growth). The correlation between CD34+ cell and CFU-GM counts was evaluated by linear regression analysis.
Results
The CB units were stored during 11-20 years (mean 16±3). We analyzed 12 CB because 3 bags were broken during storage (20%); this percentage is much higher than that found in our daily practice (4%), which may be caused by the long cryopreservation period. Visual examination of the product showed evidence of hemolysis in 10 CB and fibrin clots in 1.The recovery of total nucleated cells (TNC) was 73±14%; CD34+ cells enumeration 48.3±29.3x105 and CFU-GM quantification 28.5±19.3x104. The TNC and CD34+ cells viability was 70±8% and 91±5%, respectively. We obtained a strong positive relationship between CD34+ cell and CFU-GM (R2=0.758). We found 4 contaminated CB, one of them was already positive after processing (Enterococcus faecalis, Escherichia coli and Streptococcus mitis). We hypothesize that the pre-freezing hemocultures were not representative of the product; however, we cannot exclude a cross-contamination during the storage.
Conclusion
Despite our small serie, the results of cellular viability, purity and potency indicate that long-term cryopreservation does not negatively affect the quality of CB units for further use, even in the presence of contamination, hemolysis signs and aggregates. We think that every cord blood bank should have an expert to help transplant physicians select the best cord blood for his patient based on the control quality results performed before final release.
Session topic: E-poster
Keyword(s): Cord blood CD34+ Cells, Cryopreservation
Type: Publication Only
Background
Cord Blood (CB) is a well-recognized source of stem cells for hematopoietic transplant. Its main advantage is to be stored for long-term with a ready availability for the intended recipient and no risk for the donor.
Aims
The aim of this study was to evaluate the quality of 15 cord blood units cryopreserved in our direct donation bank. for the intended recipient and no risk for the donor.
Methods
All these units were processed without red cells reduction, in a 24 hours period after collection, then were cryopreserved with a controlled-rate freezer and transferred to a liquid nitrogen container. We used cryogenic bags (Kapton/Teflon), highly resistant to very low temperatures. However, due to missing data we do not considered these CB units suitable for transplant. In this study, the CB were thawed and washed based on the New York Blood Center method in our ISO 7 cleanroom, inside of an air flow cabinet. We performed cellular counts, viability assays (by flow cytometry using 7-aminoactinomycin), sterility testing (bacterial and fungal cultures) and clonogenicity (colony-forming units-granulocyte/macrophage - CFU-GM - growth). The correlation between CD34+ cell and CFU-GM counts was evaluated by linear regression analysis.
Results
The CB units were stored during 11-20 years (mean 16±3). We analyzed 12 CB because 3 bags were broken during storage (20%); this percentage is much higher than that found in our daily practice (4%), which may be caused by the long cryopreservation period. Visual examination of the product showed evidence of hemolysis in 10 CB and fibrin clots in 1.The recovery of total nucleated cells (TNC) was 73±14%; CD34+ cells enumeration 48.3±29.3x105 and CFU-GM quantification 28.5±19.3x104. The TNC and CD34+ cells viability was 70±8% and 91±5%, respectively. We obtained a strong positive relationship between CD34+ cell and CFU-GM (R2=0.758). We found 4 contaminated CB, one of them was already positive after processing (Enterococcus faecalis, Escherichia coli and Streptococcus mitis). We hypothesize that the pre-freezing hemocultures were not representative of the product; however, we cannot exclude a cross-contamination during the storage.
Conclusion
Despite our small serie, the results of cellular viability, purity and potency indicate that long-term cryopreservation does not negatively affect the quality of CB units for further use, even in the presence of contamination, hemolysis signs and aggregates. We think that every cord blood bank should have an expert to help transplant physicians select the best cord blood for his patient based on the control quality results performed before final release.
Session topic: E-poster
Keyword(s): Cord blood CD34+ Cells, Cryopreservation
Abstract: PB2187
Type: Publication Only
Background
Cord Blood (CB) is a well-recognized source of stem cells for hematopoietic transplant. Its main advantage is to be stored for long-term with a ready availability for the intended recipient and no risk for the donor.
Aims
The aim of this study was to evaluate the quality of 15 cord blood units cryopreserved in our direct donation bank. for the intended recipient and no risk for the donor.
Methods
All these units were processed without red cells reduction, in a 24 hours period after collection, then were cryopreserved with a controlled-rate freezer and transferred to a liquid nitrogen container. We used cryogenic bags (Kapton/Teflon), highly resistant to very low temperatures. However, due to missing data we do not considered these CB units suitable for transplant. In this study, the CB were thawed and washed based on the New York Blood Center method in our ISO 7 cleanroom, inside of an air flow cabinet. We performed cellular counts, viability assays (by flow cytometry using 7-aminoactinomycin), sterility testing (bacterial and fungal cultures) and clonogenicity (colony-forming units-granulocyte/macrophage - CFU-GM - growth). The correlation between CD34+ cell and CFU-GM counts was evaluated by linear regression analysis.
Results
The CB units were stored during 11-20 years (mean 16±3). We analyzed 12 CB because 3 bags were broken during storage (20%); this percentage is much higher than that found in our daily practice (4%), which may be caused by the long cryopreservation period. Visual examination of the product showed evidence of hemolysis in 10 CB and fibrin clots in 1.The recovery of total nucleated cells (TNC) was 73±14%; CD34+ cells enumeration 48.3±29.3x105 and CFU-GM quantification 28.5±19.3x104. The TNC and CD34+ cells viability was 70±8% and 91±5%, respectively. We obtained a strong positive relationship between CD34+ cell and CFU-GM (R2=0.758). We found 4 contaminated CB, one of them was already positive after processing (Enterococcus faecalis, Escherichia coli and Streptococcus mitis). We hypothesize that the pre-freezing hemocultures were not representative of the product; however, we cannot exclude a cross-contamination during the storage.
Conclusion
Despite our small serie, the results of cellular viability, purity and potency indicate that long-term cryopreservation does not negatively affect the quality of CB units for further use, even in the presence of contamination, hemolysis signs and aggregates. We think that every cord blood bank should have an expert to help transplant physicians select the best cord blood for his patient based on the control quality results performed before final release.
Session topic: E-poster
Keyword(s): Cord blood CD34+ Cells, Cryopreservation
Type: Publication Only
Background
Cord Blood (CB) is a well-recognized source of stem cells for hematopoietic transplant. Its main advantage is to be stored for long-term with a ready availability for the intended recipient and no risk for the donor.
Aims
The aim of this study was to evaluate the quality of 15 cord blood units cryopreserved in our direct donation bank. for the intended recipient and no risk for the donor.
Methods
All these units were processed without red cells reduction, in a 24 hours period after collection, then were cryopreserved with a controlled-rate freezer and transferred to a liquid nitrogen container. We used cryogenic bags (Kapton/Teflon), highly resistant to very low temperatures. However, due to missing data we do not considered these CB units suitable for transplant. In this study, the CB were thawed and washed based on the New York Blood Center method in our ISO 7 cleanroom, inside of an air flow cabinet. We performed cellular counts, viability assays (by flow cytometry using 7-aminoactinomycin), sterility testing (bacterial and fungal cultures) and clonogenicity (colony-forming units-granulocyte/macrophage - CFU-GM - growth). The correlation between CD34+ cell and CFU-GM counts was evaluated by linear regression analysis.
Results
The CB units were stored during 11-20 years (mean 16±3). We analyzed 12 CB because 3 bags were broken during storage (20%); this percentage is much higher than that found in our daily practice (4%), which may be caused by the long cryopreservation period. Visual examination of the product showed evidence of hemolysis in 10 CB and fibrin clots in 1.The recovery of total nucleated cells (TNC) was 73±14%; CD34+ cells enumeration 48.3±29.3x105 and CFU-GM quantification 28.5±19.3x104. The TNC and CD34+ cells viability was 70±8% and 91±5%, respectively. We obtained a strong positive relationship between CD34+ cell and CFU-GM (R2=0.758). We found 4 contaminated CB, one of them was already positive after processing (Enterococcus faecalis, Escherichia coli and Streptococcus mitis). We hypothesize that the pre-freezing hemocultures were not representative of the product; however, we cannot exclude a cross-contamination during the storage.
Conclusion
Despite our small serie, the results of cellular viability, purity and potency indicate that long-term cryopreservation does not negatively affect the quality of CB units for further use, even in the presence of contamination, hemolysis signs and aggregates. We think that every cord blood bank should have an expert to help transplant physicians select the best cord blood for his patient based on the control quality results performed before final release.
Session topic: E-poster
Keyword(s): Cord blood CD34+ Cells, Cryopreservation
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