TransFix® is designed for stabilisation of peripheral blood and cerebrospinal fluid to enumerate lymphocyte subpopulations by flow cytometry. However, researchers have developed many new and innovative uses of TransFix.
We’ve presented a selection of publications to highlight the versatility of TransFix below. Read on to find out the different ways that researchers benefit from sample stabilisation.
If you have a problem with sample degradation in your research, contact us to see if TransFix could help.
Extraction of mRNA from Samples
It is well documented that TransFix treated blood is compatible with isolation of circulating tumour cells, but Baird et al. describes a novel method of isolating CTCs, and the first published method of extracting mRNA from TransFix-treated samples for RT-qPCR analysis.
- Baird et al. (2019) Enumeration of rare cells in whole blood by signal ion emission reactive release amplification (SIERRA) with same-sample RNA analysis. Anal. Chem 91(3); 2028-2034. https://doi.org/10.1021/acs.analchem.8b04446
Compatible with Multiple Anticoagulants
Cytomark have validated TransFix for use with EDTA anticoagulant for lymphocyte immunophenotyping, however Olivares et al. collect samples on sodium heparin tubes showing compatibility with other anticoagulants in the context of a clinical trial.
- Olivares et al. (2014) Double-blind, randomised, placebo-controlled intervention trial to evaluate the effects of Bifidobacterium longum CECT 7347 in children with newly diagnosed coeliac disease. British Journal of Nutrition 112(1); 30-40. https://doi.org/10.1017/S0007114514000609
Stabilisation of 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 Progenitor Cells which have been referenced by multiple papers.
- Farinacci et al. (2018) Circulating endothelial cells as biomarker for cardiovascular diseases. Res Pract Thromb Haemost. 2018; 1–10. https://doi.org/10.1002/rth2.12158
- Ribeiro et al. (2017) Effects of resistance exercise on endothelial progenitor cell mobilization in women. SCIENTIFIC Reports 7; 17880. https://doi.org/10.1038/s41598-017-18156-6
- 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
- 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
Stabilisation of T-Cell Activation Markers
Measurement of T-cell activation can be important when assessing immune responses, however cells can become artificially activated over time once removed from the body.
TransFix has been used to stabilise activated T-cell populations. Almost 10 years ago Leligdowicz et al. used TransFix to assess the relationship between HIV-2 plasma virus load and immune system activation using HLA-DR and CD38. Since then other publications include Olivares et al. who identified activated CD45+HLA-DR+ positive T-cells from TransFix-treated blood in relation to coeliac disease, and Poirier et al. used TransFix to stabilise the immune profile in their clinical trial, using CD69+ and CD25+ to identify activated T-cells in response to anti-CD28 therapy
- Leligdowicz et al. (2010) Direct Relationship between Virus Load and Systemic Immune Activation in HIV-2 Infection. J Infect Dis 201(1); 114-122. https://doi.org/10.1086/648733
- Olivares et al. (2014) Double Blind, randomised, placebo-controlled intervention trial to evaluate the effects of Bifido longum CECT 7347 in children with newly diagnosed coeliac disease. British Journal of Nutrition 112(1); 30-40. https://doi.org/10.1017/S0007114514000609
- Poirier et al. (2016) First-in-Human Study in Healthy Subjects with FR104, a Pegylated Monoclonal Antibody Fragment Antagonist of CD28. J Immunol 197(12); 4593-4602. https://doi.org/10.4049/jimmunol.1601538
Preserving Other Biological Fluids
TransFix has been shown to preserve cells in a variety of biological fluids. Noël et al. used TransFix to stabilise Bronchoalveolar Lavage Fluids (BALF) from Rats to assess pulmonary toxicity of aerosols. Another example is stabilisation of stromal vascular fraction by Karsten et al., who demonstrated the application of TransFix for biological mixed cell populations.
- Noël et al. (2012)Effects of inhaled nano-TiO2 aerosols showing two distinct agglomeration states on rat lungs. Toxicology Letters 214; 109– 119 https://doi.org/10.1016/j.toxlet.2012.08.019
- Karsten et al. (2014) Evaluation of TransFix® Mediated Stabilisation of Adipose-Derived Stromal Vascular Fraction for Delayed Flow Cytometry Analysis. Open Journal of Regenerative Medicine 3; 54-63. https://doi.org/10.4236/ojrm.2014.33007
Stem Cell Enumeration
CD34+ population enumeration has multiple applications, for example to indicate whether a stem-cell harvest is ready to be performed, or to assess B-cell precursors in ALL samples. However, rare CD34+ populations can be hard to define due to sample degradation.
There are several references where researchers analyse CD34+ populations with TransFix-treated samples:
- 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:e009444 https://doi.org/10.1161/JAHA.118.009444
- Levinsen et al. (2016) Leukemic blasts are present at low levels in spinal fluid in one-third of childhood acute lymphoblastic leukemia cases. Pediatr Blood Cancer 63(11); 1935-1942 https://doi.org/10.1002/pbc.26128
- Beiral et al. (2014) The impact of stem cells on electron fluxes, proton translocation, and ATP synthesis in kidney mitochondria after ischemia/reperfusion. Cell Transplant. 23(2); 207-20. https://doi.org/10.3727/096368912X659862
For more examples of publications referencing the use of TransFix, visit the bibliography section on the Cytomark website!
If you have any questions about the use of TransFix please contact us and we’ll be happy to help:
Cytomark is a division of Caltag Medsystems Ltd. The published applications in this email do not relate to an IVD product claim. Cytomark have not verified these findings.