The Assessment of Parameters Affecting the Quality of Cord Blood by the Appliance of the Annexin V Staining Method and Correlation with CFU Assays
Open Access
- 7 March 2013
- journal article
- research article
- Published by Hindawi Limited in Stem Cells International
- Vol. 2013, 1-10
- https://doi.org/10.1155/2013/823912
Abstract
The assessment of nonviable haematopoietic cells by Annexin V staining method in flow cytometry has recently been published by Duggleby et al. Resulting in a better correlation with the observed colony formation in methylcellulose assays than the standard ISHAGE protocol, it presents a promising method to predict cord blood potency. Herein, we applied this method for examining the parameters during processing which potentially could affect cord blood viability. We could verify that the current standards regarding time and temperature are sufficient, since no significant difference was observed within 48 hours or in storage at up to . However, the addition of DMSO for cryopreservation alone leads to an inevitable increase in nonviable haematopoietic stem cells from initially 14.8% ± 4.3% to at least 30.6% ± 5.5%. Furthermore, CFU-assays with varied seeding density were performed in order to evaluate the applicability as a quantitative method. The results revealed that only in a narrow range reproducible clonogenic efficiency (ClonE) could be assessed, giving at least a semiquantitative estimation. We conclude that both Annexin V staining method and CFU-assays with defined seeding density are reliable means leading to a better prediction of the final potency. Especially Annexin V, due to its fast readout, is a practical tool for examining and optimising specific steps in processing, while CFU-assays add a functional confirmation.Keywords
Funding Information
- Eurocord
This publication has 25 references indexed in Scilit:
- Flow cytometry assessment of apoptotic CD34+ cells by annexin V labeling may improve prediction of cord blood potency for engraftmentTransfusion, 2011
- Total Colony-Forming Units Are a Strong, Independent Predictor of Neutrophil and Platelet Engraftment after Unrelated Umbilical Cord Blood Transplantation: A Single-Center Analysis of 435 Cord Blood TransplantsTransplantation and Cellular Therapy, 2011
- Effect of graft source on unrelated donor haemopoietic stem-cell transplantation in adults with acute leukaemia: a retrospective analysisThe Lancet Oncology, 2010
- Factors influencing cord blood viability assessment before cryopreservationTransfusion, 2010
- Quality rather than quantity: the cord blood bank dilemmaBone Marrow Transplantation, 2010
- Early apoptosis in CD34+ cells as a potential heterogeneity in quality of cryopreserved umbilical cord bloodBritish Journal of Haematology, 2006
- The ISHAGE Guidelines for CD34+ Cell Determination by Flow CytometryJournal of Hematotherapy, 1996
- Hematopoietic progenitor cells are resistant to dimethyl sulfoxide toxicityTransfusion, 1994
- Dead cell discrimination with 7‐amino‐actinomcin D in combination with dual color immunofluorescence in single laser flow cytometryCytometry, 1992
- Hematopoietic Reconstitution in a Patient with Fanconi's Anemia by Means of Umbilical-Cord Blood from an HLA-Identical SiblingNew England Journal of Medicine, 1989