Extensive early apoptosis in frozen-thawed CD34-positive stem cells decreases threshold doses for haematological recovery after autologous peripheral blood progenitor cell transplantation

F. de Boer, A. M. Dräger, H. M. Pinedo, F. L. Kessler, E. van der Wall, A. R. Jonkhoff, J. van der Lelie, P. C. Huijgens, G. J. Ossenkoppele, G. J. Schuurhuis

Research output: Contribution to journalArticle

Abstract

Stem cell doses necessary for engraftment after myeloablative therapy as defined for fresh transplants vary largely. Loss of CD34+ cell quality after cryopreservation might contribute to this variation. With a new early apoptosis assay including the vital stain Syto16, together with the permeability marker 7-AAD, CD34+ cell viability in leucapheresis samples of 49 lymphoma patients receiving a BEAM regimen was analysed. After freeze-thawing large numbers of non-viable, early apoptotic cells appeared, leading to only 42% viability compared to 72% using 7-AAD only. Based on this Syto16 staining in the frozen-thawed grafts, threshold numbers for adequate haematological recovery of 2.8-3.0 × 106 CD34+ cells/kg body weight determined for fresh grafts, now decreased to 1.2-1.3 × 106 CD34+ cells/kg. In whole blood transplantation of lymphoma patients (n = 45) receiving a BEAM-like regimen, low doses of CD34+ cells were sufficient for recovery (0.3-0.4 × 106 CD34+ cells/kg). In contrast to freeze-thawing of leucapheresis material, a high viability of CD34+ cells was preserved during storage for 3 days at 4°C, leaving threshold doses for recovery unchanged. In conclusion, the Syto16 assay reveals the presence of many more non-functional stem cells in frozen-thawed transplants than presumed thus far. This led to a factor 2.3-fold adjustment downward of viable CD34+ threshold doses for haematological recovery.

Original languageEnglish (US)
Pages (from-to)249-255
Number of pages7
JournalBone Marrow Transplantation
Volume29
Issue number3
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Cell Transplantation
Blood Cells
Stem Cells
Apoptosis
Transplants
Lymphoma
Cell Survival
Cryopreservation
Permeability
Coloring Agents
Transplantation
Body Weight
Staining and Labeling
BEAM regimen

Keywords

  • Apoptosis
  • CD34
  • Stem cell transplant
  • Syto16

ASJC Scopus subject areas

  • Hematology
  • Transplantation

Cite this

Extensive early apoptosis in frozen-thawed CD34-positive stem cells decreases threshold doses for haematological recovery after autologous peripheral blood progenitor cell transplantation. / de Boer, F.; Dräger, A. M.; Pinedo, H. M.; Kessler, F. L.; van der Wall, E.; Jonkhoff, A. R.; van der Lelie, J.; Huijgens, P. C.; Ossenkoppele, G. J.; Schuurhuis, G. J.

In: Bone Marrow Transplantation, Vol. 29, No. 3, 2002, p. 249-255.

Research output: Contribution to journalArticle

de Boer, F, Dräger, AM, Pinedo, HM, Kessler, FL, van der Wall, E, Jonkhoff, AR, van der Lelie, J, Huijgens, PC, Ossenkoppele, GJ & Schuurhuis, GJ 2002, 'Extensive early apoptosis in frozen-thawed CD34-positive stem cells decreases threshold doses for haematological recovery after autologous peripheral blood progenitor cell transplantation', Bone Marrow Transplantation, vol. 29, no. 3, pp. 249-255. https://doi.org/10.1038/sj/bmt/1703357
de Boer, F. ; Dräger, A. M. ; Pinedo, H. M. ; Kessler, F. L. ; van der Wall, E. ; Jonkhoff, A. R. ; van der Lelie, J. ; Huijgens, P. C. ; Ossenkoppele, G. J. ; Schuurhuis, G. J. / Extensive early apoptosis in frozen-thawed CD34-positive stem cells decreases threshold doses for haematological recovery after autologous peripheral blood progenitor cell transplantation. In: Bone Marrow Transplantation. 2002 ; Vol. 29, No. 3. pp. 249-255.
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abstract = "Stem cell doses necessary for engraftment after myeloablative therapy as defined for fresh transplants vary largely. Loss of CD34+ cell quality after cryopreservation might contribute to this variation. With a new early apoptosis assay including the vital stain Syto16, together with the permeability marker 7-AAD, CD34+ cell viability in leucapheresis samples of 49 lymphoma patients receiving a BEAM regimen was analysed. After freeze-thawing large numbers of non-viable, early apoptotic cells appeared, leading to only 42{\%} viability compared to 72{\%} using 7-AAD only. Based on this Syto16 staining in the frozen-thawed grafts, threshold numbers for adequate haematological recovery of 2.8-3.0 × 106 CD34+ cells/kg body weight determined for fresh grafts, now decreased to 1.2-1.3 × 106 CD34+ cells/kg. In whole blood transplantation of lymphoma patients (n = 45) receiving a BEAM-like regimen, low doses of CD34+ cells were sufficient for recovery (0.3-0.4 × 106 CD34+ cells/kg). In contrast to freeze-thawing of leucapheresis material, a high viability of CD34+ cells was preserved during storage for 3 days at 4°C, leaving threshold doses for recovery unchanged. In conclusion, the Syto16 assay reveals the presence of many more non-functional stem cells in frozen-thawed transplants than presumed thus far. This led to a factor 2.3-fold adjustment downward of viable CD34+ threshold doses for haematological recovery.",
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AU - de Boer, F.

AU - Dräger, A. M.

AU - Pinedo, H. M.

AU - Kessler, F. L.

AU - van der Wall, E.

AU - Jonkhoff, A. R.

AU - van der Lelie, J.

AU - Huijgens, P. C.

AU - Ossenkoppele, G. J.

AU - Schuurhuis, G. J.

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N2 - Stem cell doses necessary for engraftment after myeloablative therapy as defined for fresh transplants vary largely. Loss of CD34+ cell quality after cryopreservation might contribute to this variation. With a new early apoptosis assay including the vital stain Syto16, together with the permeability marker 7-AAD, CD34+ cell viability in leucapheresis samples of 49 lymphoma patients receiving a BEAM regimen was analysed. After freeze-thawing large numbers of non-viable, early apoptotic cells appeared, leading to only 42% viability compared to 72% using 7-AAD only. Based on this Syto16 staining in the frozen-thawed grafts, threshold numbers for adequate haematological recovery of 2.8-3.0 × 106 CD34+ cells/kg body weight determined for fresh grafts, now decreased to 1.2-1.3 × 106 CD34+ cells/kg. In whole blood transplantation of lymphoma patients (n = 45) receiving a BEAM-like regimen, low doses of CD34+ cells were sufficient for recovery (0.3-0.4 × 106 CD34+ cells/kg). In contrast to freeze-thawing of leucapheresis material, a high viability of CD34+ cells was preserved during storage for 3 days at 4°C, leaving threshold doses for recovery unchanged. In conclusion, the Syto16 assay reveals the presence of many more non-functional stem cells in frozen-thawed transplants than presumed thus far. This led to a factor 2.3-fold adjustment downward of viable CD34+ threshold doses for haematological recovery.

AB - Stem cell doses necessary for engraftment after myeloablative therapy as defined for fresh transplants vary largely. Loss of CD34+ cell quality after cryopreservation might contribute to this variation. With a new early apoptosis assay including the vital stain Syto16, together with the permeability marker 7-AAD, CD34+ cell viability in leucapheresis samples of 49 lymphoma patients receiving a BEAM regimen was analysed. After freeze-thawing large numbers of non-viable, early apoptotic cells appeared, leading to only 42% viability compared to 72% using 7-AAD only. Based on this Syto16 staining in the frozen-thawed grafts, threshold numbers for adequate haematological recovery of 2.8-3.0 × 106 CD34+ cells/kg body weight determined for fresh grafts, now decreased to 1.2-1.3 × 106 CD34+ cells/kg. In whole blood transplantation of lymphoma patients (n = 45) receiving a BEAM-like regimen, low doses of CD34+ cells were sufficient for recovery (0.3-0.4 × 106 CD34+ cells/kg). In contrast to freeze-thawing of leucapheresis material, a high viability of CD34+ cells was preserved during storage for 3 days at 4°C, leaving threshold doses for recovery unchanged. In conclusion, the Syto16 assay reveals the presence of many more non-functional stem cells in frozen-thawed transplants than presumed thus far. This led to a factor 2.3-fold adjustment downward of viable CD34+ threshold doses for haematological recovery.

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