Viability of cryopreserved BM progenitor cells stored for more than a decade

Albert D. Donnenberg, E. K. Koch, D. L. Griffin, H. M. Stanczak, J. E. Kiss, T. M. Carlos, D. M. BuchBarker, Andrew M Yeager

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Background: PBPC or BM is increasingly being harvested in remission for possible use in the event of relapse. Although the value of this approach has not been demonstrated, the long-term storage of progenitor cell components has become commonplace in many facilities. Methods: We used multi-parameter flow cytometry to determine the viability of 11 long-term cryopreserved BM components (mean = 11.8 years) in liquid phase nitrogen. The components, prepared for autotransplantation but deaccessioned after confirming patient death, were carefully thawed, washed, and assayed immediately. The flow cytometry assay was performed according to the ISHAGE protocol, modified by the addition of 7AAD for analysis of progenitor viability (CD45+ CD34+ 7AAD-) and total leukocyte viability (CD45+ 7AAD-). In addition, total viability was assessed by fluorescence microscopy using acridine orange dye exclusion, granulocyte-monocyte colony-forming units (CFU-GM) were measured after 14 days culture. Results: Leukocyte viability by flow cytometry and fluorescence microscopy agreed well (r2 = 0.55, slope = 0.83, P < 0.0005 by linear regression). CFU-GM did not correlate with CD34% or any of the viability parameters. Compared with short-term stored (mean = 33 days) PBPC assayed at infusion, long-term stored BM had a comparable percentage of CD34+ cells, comparable CFU-GM activity, increased CD34 viability, but decreased total cell viability, the latter most likely due to an increased proportion of differentiated myeloid cells. Discussion: The results indicate that BM products can be cryopreserved for more than a decade without apparent loss of progenitor activity, as measured by these laboratory surrogates. This agrees with clinical anecdotes describing successful engraftment with long-term stored BM, and argues that expiration dates cannot be set for cryopreserved hematopoietic stem-cell components stored in liquid phase nitrogen.

Original languageEnglish (US)
Pages (from-to)157-163
Number of pages7
JournalCytotherapy
Volume4
Issue number2
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Granulocyte-Macrophage Progenitor Cells
Flow Cytometry
Stem Cells
Cellular Structures
Fluorescence Microscopy
Leukocytes
Nitrogen
Anecdotes
Acridine Orange
Autologous Transplantation
Myeloid Cells
Hematopoietic Stem Cells
Granulocytes
Monocytes
Linear Models
Cell Survival
Coloring Agents
Recurrence

Keywords

  • Bone marrow
  • CD34 enumeration
  • Cryopreservation
  • Flow cytometry
  • Peripheral blood progenitor cells
  • Viability

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy
  • Pharmacology

Cite this

Donnenberg, A. D., Koch, E. K., Griffin, D. L., Stanczak, H. M., Kiss, J. E., Carlos, T. M., ... Yeager, A. M. (2002). Viability of cryopreserved BM progenitor cells stored for more than a decade. Cytotherapy, 4(2), 157-163. https://doi.org/10.1080/146532402317381866

Viability of cryopreserved BM progenitor cells stored for more than a decade. / Donnenberg, Albert D.; Koch, E. K.; Griffin, D. L.; Stanczak, H. M.; Kiss, J. E.; Carlos, T. M.; BuchBarker, D. M.; Yeager, Andrew M.

In: Cytotherapy, Vol. 4, No. 2, 2002, p. 157-163.

Research output: Contribution to journalArticle

Donnenberg, AD, Koch, EK, Griffin, DL, Stanczak, HM, Kiss, JE, Carlos, TM, BuchBarker, DM & Yeager, AM 2002, 'Viability of cryopreserved BM progenitor cells stored for more than a decade', Cytotherapy, vol. 4, no. 2, pp. 157-163. https://doi.org/10.1080/146532402317381866
Donnenberg AD, Koch EK, Griffin DL, Stanczak HM, Kiss JE, Carlos TM et al. Viability of cryopreserved BM progenitor cells stored for more than a decade. Cytotherapy. 2002;4(2):157-163. https://doi.org/10.1080/146532402317381866
Donnenberg, Albert D. ; Koch, E. K. ; Griffin, D. L. ; Stanczak, H. M. ; Kiss, J. E. ; Carlos, T. M. ; BuchBarker, D. M. ; Yeager, Andrew M. / Viability of cryopreserved BM progenitor cells stored for more than a decade. In: Cytotherapy. 2002 ; Vol. 4, No. 2. pp. 157-163.
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abstract = "Background: PBPC or BM is increasingly being harvested in remission for possible use in the event of relapse. Although the value of this approach has not been demonstrated, the long-term storage of progenitor cell components has become commonplace in many facilities. Methods: We used multi-parameter flow cytometry to determine the viability of 11 long-term cryopreserved BM components (mean = 11.8 years) in liquid phase nitrogen. The components, prepared for autotransplantation but deaccessioned after confirming patient death, were carefully thawed, washed, and assayed immediately. The flow cytometry assay was performed according to the ISHAGE protocol, modified by the addition of 7AAD for analysis of progenitor viability (CD45+ CD34+ 7AAD-) and total leukocyte viability (CD45+ 7AAD-). In addition, total viability was assessed by fluorescence microscopy using acridine orange dye exclusion, granulocyte-monocyte colony-forming units (CFU-GM) were measured after 14 days culture. Results: Leukocyte viability by flow cytometry and fluorescence microscopy agreed well (r2 = 0.55, slope = 0.83, P < 0.0005 by linear regression). CFU-GM did not correlate with CD34{\%} or any of the viability parameters. Compared with short-term stored (mean = 33 days) PBPC assayed at infusion, long-term stored BM had a comparable percentage of CD34+ cells, comparable CFU-GM activity, increased CD34 viability, but decreased total cell viability, the latter most likely due to an increased proportion of differentiated myeloid cells. Discussion: The results indicate that BM products can be cryopreserved for more than a decade without apparent loss of progenitor activity, as measured by these laboratory surrogates. This agrees with clinical anecdotes describing successful engraftment with long-term stored BM, and argues that expiration dates cannot be set for cryopreserved hematopoietic stem-cell components stored in liquid phase nitrogen.",
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T1 - Viability of cryopreserved BM progenitor cells stored for more than a decade

AU - Donnenberg, Albert D.

AU - Koch, E. K.

AU - Griffin, D. L.

AU - Stanczak, H. M.

AU - Kiss, J. E.

AU - Carlos, T. M.

AU - BuchBarker, D. M.

AU - Yeager, Andrew M

PY - 2002

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N2 - Background: PBPC or BM is increasingly being harvested in remission for possible use in the event of relapse. Although the value of this approach has not been demonstrated, the long-term storage of progenitor cell components has become commonplace in many facilities. Methods: We used multi-parameter flow cytometry to determine the viability of 11 long-term cryopreserved BM components (mean = 11.8 years) in liquid phase nitrogen. The components, prepared for autotransplantation but deaccessioned after confirming patient death, were carefully thawed, washed, and assayed immediately. The flow cytometry assay was performed according to the ISHAGE protocol, modified by the addition of 7AAD for analysis of progenitor viability (CD45+ CD34+ 7AAD-) and total leukocyte viability (CD45+ 7AAD-). In addition, total viability was assessed by fluorescence microscopy using acridine orange dye exclusion, granulocyte-monocyte colony-forming units (CFU-GM) were measured after 14 days culture. Results: Leukocyte viability by flow cytometry and fluorescence microscopy agreed well (r2 = 0.55, slope = 0.83, P < 0.0005 by linear regression). CFU-GM did not correlate with CD34% or any of the viability parameters. Compared with short-term stored (mean = 33 days) PBPC assayed at infusion, long-term stored BM had a comparable percentage of CD34+ cells, comparable CFU-GM activity, increased CD34 viability, but decreased total cell viability, the latter most likely due to an increased proportion of differentiated myeloid cells. Discussion: The results indicate that BM products can be cryopreserved for more than a decade without apparent loss of progenitor activity, as measured by these laboratory surrogates. This agrees with clinical anecdotes describing successful engraftment with long-term stored BM, and argues that expiration dates cannot be set for cryopreserved hematopoietic stem-cell components stored in liquid phase nitrogen.

AB - Background: PBPC or BM is increasingly being harvested in remission for possible use in the event of relapse. Although the value of this approach has not been demonstrated, the long-term storage of progenitor cell components has become commonplace in many facilities. Methods: We used multi-parameter flow cytometry to determine the viability of 11 long-term cryopreserved BM components (mean = 11.8 years) in liquid phase nitrogen. The components, prepared for autotransplantation but deaccessioned after confirming patient death, were carefully thawed, washed, and assayed immediately. The flow cytometry assay was performed according to the ISHAGE protocol, modified by the addition of 7AAD for analysis of progenitor viability (CD45+ CD34+ 7AAD-) and total leukocyte viability (CD45+ 7AAD-). In addition, total viability was assessed by fluorescence microscopy using acridine orange dye exclusion, granulocyte-monocyte colony-forming units (CFU-GM) were measured after 14 days culture. Results: Leukocyte viability by flow cytometry and fluorescence microscopy agreed well (r2 = 0.55, slope = 0.83, P < 0.0005 by linear regression). CFU-GM did not correlate with CD34% or any of the viability parameters. Compared with short-term stored (mean = 33 days) PBPC assayed at infusion, long-term stored BM had a comparable percentage of CD34+ cells, comparable CFU-GM activity, increased CD34 viability, but decreased total cell viability, the latter most likely due to an increased proportion of differentiated myeloid cells. Discussion: The results indicate that BM products can be cryopreserved for more than a decade without apparent loss of progenitor activity, as measured by these laboratory surrogates. This agrees with clinical anecdotes describing successful engraftment with long-term stored BM, and argues that expiration dates cannot be set for cryopreserved hematopoietic stem-cell components stored in liquid phase nitrogen.

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KW - CD34 enumeration

KW - Cryopreservation

KW - Flow cytometry

KW - Peripheral blood progenitor cells

KW - Viability

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