Large populations of non-clonogenic early apoptotic CD34-positive cells are present in frozen-thawed peripheral blood stem cell transplants

Bone Marrow Transplantation
Gerrit Jan SchuurhuisHenk J Broxterman

Abstract

Apoptosis is the common cell death pathway which is initiated by a variety of different stimuli. The recognition of early apoptotic events would markedly improve reliability and convenience of apoptosis assays. In the present study the vital stain SytoR 16 in combination with the permeability marker 7-amino actinomycin D, (7-AAD) has been used to identify an early stage of apoptosis, not detected with trypan blue or 7-AAD alone or with conventional apoptosis tests and not consistently and only partly detected by the early apoptosis marker annexin V. The method was established using solid tumour cell lines treated with TNF. Subsequently we applied it to determine apoptotic populations in CD34(+) peripheral blood progenitor cells obtained from growth factor and/or chemotherapy mobilised patients and frozen/thawed according to standard stem cell transplantation protocols. In a cell line model as well as CD34(+) progenitor cells, different subpopulations with decreased SytoR 16 fluorescence (SytoR 16int or SytoR 16low, compared with the normal SytoR 16high) appeared which are not, or only partly, apoptotic using conventional techniques including morphology or 7-AAD staining: eg percentages of SytoR 16(int)/7-AAD(-) and SytoR 16(low...Continue Reading

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Citations

Jul 14, 2007·Biotechnology Letters·Hyun-Seok KimHyo-Il Jung
Jan 19, 2002·Journal of Hematotherapy & Stem Cell Research·A M de KreukA R Jonkhoff
Feb 20, 2003·Journal of Hematotherapy & Stem Cell Research·Fransien de BoerGerrit J Schuurhuis
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Nov 1, 2004·Molecular Plant Pathology·Peter VAN BaarlenJan A L VAN Kan
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May 22, 2008·British Journal of Haematology·Ilse HoutenbosArjan A van de Loosdrecht
Jan 4, 2007·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Donald WlodkowicJukka Pelkonen
Nov 15, 2013·Transfusion·Martin G GuttridgeSuzanne M Watt
Sep 17, 2005·Journal of Immunological Methods·Rosemary L Sparrow, Emma Tippett
Jan 5, 2010·Organogenesis·John G BaustJohn M Baust
Mar 1, 2009·Biopreservation and Biobanking·John M BaustJohn G Baust
Nov 7, 2012·Cell Transplantation·Pei-Ming ChuShih-Hwa Chiou
Mar 9, 2018·Transfusion·Sofia FrändbergLars Palmqvist
Aug 30, 2018·Stem Cells·Sehar AfreenMiriam Erlacher

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