Stabilisation of Circulating Endothelial Progenitor Cells

TransFix stabilises lymphocytes for up to 14 days, but other cell populations have also been shown to be stable with TransFix. One example is Circulating Endothelial and Endothelial Progenitor Cells which have been referenced by multiple papers:

TransFix for Delayed Flow Cytometry of Endothelial Progenitor Cells

Here, Hoymans et al. report results for the flow cytometric enumeration of circulating endothelial progenitor cells (EPC) and angiogenic T cells using TransFix-treated whole blood obtained from healthy adults and those with cardiovascular disease. Both cell types promote neovascularization and vascular homeostasis, and are therefore of interest as potential biomarkers for cardiovascular disease. The authors report that with the addition of TransFix to whole blood, analysis can be postponed up to 7 days after blood collection, facilitating laboratory workflow, as well as the organisation of multicenter studies, which requires analyses to be conducted in a central core laboratory.

TransFix-Enabled Transport of Endothelial Progenitor Cells and Circulating Endothelial Cells

The aim of this study was to determine differences in the number of endothelial progenitor cells and/or circulating endothelial cells found in peripheral blood in patients treated with either ticagrelor or clopidogrel during non-ST-segment-elevation myocardial infarction. In this multicentre, randomized study (NCT02244710), all samples were kept in TransFix to enable adequate shipping to a central laboratory for analysis within 5 days Markers included: CD34, CD133, CD45, CD146, CD31, KDR (CD309), 7AAD, and Syto-16. Generic 7AAD and Syto-16 markers were used to define cell viability and nuclear cells, respectively.

  • Diego-Nieto et al. (2018) No Differences in Levels of Circulating Progenitor Endothelial Cells or Circulating Endothelial Cells Among Patients Treated With Ticagrelor Compared With Clopidogrel During Non-ST-Segment-Elevation Myocardial Infarction. Journal of the American Heart Association. 7(19): e009444 https://doi.org/10.1161/JAHA.118.009444

Preserving Circulating Endothelial Cells as Biomarker for Cardiovascular Diseases

Farinacci et al.’s 2018 paper document the validation of a new assay to evaluate Circulating Endothelial Cells (CEC) and endothelial progenitor cells (EPCs) as a biomarker for cardiovascular diseases. All blood samples were collected into TransFix TVT’s (3ml) and analysed by FACS. CECs were defined as DNA+, CD45dim, CD31+, CD146+ and CD36+/-.
  • Farinacci et al. (2018) Circulating endothelial cells as biomarker for cardiovascular diseases. Res Pract Thromb Haemost. 3(1): 49-58. https://doi.org/10.1002/rth2.12158

Analysing Stabilised Circulating Endothelial Cells in Response to Exercise

In 2017, Ribeiro et al. studied the effect of a single bout of resistance exercise at different intensities on the mobilization of circulating endothelial progenitor cells (EPCs) over 24 hours in women. The Authors used TransFix to stabilise blood collected on EDTA until they could analyse by flow cytometry 3-4 days after collection (CD34, CD309, CD45). They note in the paper that it was possible to analyse TransFix stabilized blood cells (EPCs) up to seven days after blood collection

TransFix Stabilisation for the Detection and Enumeration of Rare Circulating Populations

In this study, Magbauna et al. evaluate the performance of a novel approach for detection and enumeration of multiple rare cell populations of metastatic breast and lung cancer patients using an automated microfluidic filtration and multiplex immunoassay strategy. Different circulating rare cell populations were detected and enumerated, including circulating tumour cells (CTCs), circulating mesenchymal cells (CMCs), circulating endothelial cells (CECs), and putative circulating stem cells (CSCs). Simultaneous assessment of CTCs, CMCs, CSCs and CECs may provide new tools to study mechanisms of disease progression and treatment response/resistance. The highly controlled filtration process and the multi-step staining parameters were optimised to minimise the detection of false positives in healthy donor blood. Use of TransFix®, along with controlled shipping and storage conditions contributed to the high rate of reportable results (98%). Blood was collected into tubes containing K3EDTA and 0.45mL TransFix® in customised TransFix/EDTA Vacuum Blood Collection Tubes (CTC-TVT-09-50).

  • Magbanua et al. (2015) A Novel Strategy for Detection and Enumeration of Circulating Rare Cell Populations in Metastatic Cancer Patients Using Automated Microfluidic Filtration and Multiplex Immunoassay. PLoS ONE 10(10); e0141166. https://doi.org/10.1371/journal.pone.0141166

Sample Collection Considerations for Dim Endothelial Progenitor Cells

A comprehensive review of characterisation of circulating endothelial progenitor cells with a particular focus on the considerations surrounding enumeration by flow cytometry and their impact on cardiovascular disease. The authors note TransFix can be used to prolong sample storage time for 7 days for this type of analysis.

  • Van Craenenbroeck et al. (2013) Quantification of circulating CD34+/KDR+/CD45 dim endothelial progenitor cells: analytical considerations. International Journal of Cardiology 167(5); 1688-1695. https://doi.org/10.1016/j.ijcard.2012.10.047

For more examples of publications referencing the use of TransFix, visit our bibliography section!

View Bibliography

Please note that the published applications in this blog do not relate to an IVD product claim. Cytomark have not verified these findings.

TransFix Products

TransFix® has been validated with a number of specific sample types and optimised with specially designed sample collection tubes:

If you have a problem with sample degradation in your research, contact us to see if TransFix could help.

About the Author

Cytomark