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Bibliography

2014

Human Blood Stabilisation - Other

Enumeration of circulating fibrocytes for clinical use is asthma by oprimized single-platform flow cytometry assay

Sample Type: Circulating fibrocytes

Analysis Method: Flow Cytometry

Authors

L Bianchetti, M Isgro, MA Marini, M Schmidt

Publication

BBA Clinical, Volume 1, June 2014, Pages 52–58

Elevated numbers of circulating fibrocytes is emerging as an easily accessible biomarker for asthma. In this report, blood samples were taken from 44 patients with asthma of various severity and 14 age-matched healthy individuals. Fibrocytes were quantified using flow cytometry. Blood samples were either processed within four hours or mixed with the cellular surface antigen-stabilizing agent TransFix and stored at 4 degrees Celsius for up to 96 hours before processing. Robust intra-assay variability demonstrated a good agreement between the fibrocyte counts measured in fresh whole blood and the fibrocyte counts obtained in stored whole blood at 48 and 96h.

http://www.sciencedirect.com/science/article/pii/S2214647414000099

Evaluation of TransFix® mediated stabilisation of adipose-derived stromal vascular fraction for delayed flow cytometry analysis

Sample Type: Adipose-derived stromal vascular fraction

Analysis Method: Flow Cytometry

Authors

Karsten,E., Sung,J., Morgan,C., Herbert,B., Vesey,G.

Publication

Open Journal of Regenerative Medicine (2014), 3, 54-63

In multi-centre studies where stromal vascular fraction (SVF) is used for immediate treatment, post-hoc assessment at a central or core facility of cell numbers, cell viability and immunophenotype may be necessary. Additional time taken to ship and store these samples can lead to time-dependant loss of cell markers and a reduction in cell viability. This paper provides an assessment of the suitability of TransFix for the purpose of SVF stabilisation across 77 consenting patients. The data shows that following treatment with TransFix the samples remain stable for up to seven days providing accurate total nucleated cell counts, stable populations of CD90, CD31 and CD45 markers and the potential to back calculate cell viability in the original sample. The authors also report an apparent increase in cell numbers post-stabilisation that is hypothesised to be due to a reduction in cell clumping as a result of treatment with TransFix.

http://www.scirp.org/journal/PaperInformation.aspx?paperID=49498#.VLPTzCusWTI

The role of tissue factor in activation coagulation system at pregnancy complications

Sample Type: Monocytes

Analysis Method: Flow Cytometry

Authors

M. Prochazka, L. Slavik L, J. Prochazkova, J. Ulehlova, M. Novák, P. Polak, R. Pilka

Publication

Thrombosis Research, 133: S109

The authors of this poster abstract propose a model to monitor activation of the coagulation system in preeclampsia and other pregnancy complications with Tissue Factor (TF) expression on monocytes by ?ow cytometry, and simultaneously ?xing the TF-induced thrombin generation in plasma. To determine expression of tissue factor (CD142) on monocytes, the authors used multicolour ?ow cytometry (CD45, CD14, CD16b, and CD142), and used TransFix to stabilise samples that could not be analysed within 2 hours.

https://www.mendeley.com/research/role-tissue-factor-activation-coagulation-system-pregnancy-complications/

CSF Stabilisation

Contribution of cerebrospinal fluid sCD19 levels to the detection of lymphoma and its impact on disease outcome

Analysis Method: Flow Cytometry

Authors

C. Muniz, L. Martin-Martin, A. Lopez

Publication

Blood, 123:1864-1869; doi: https://doi.org/10.1182/blood-2013-11-537993

The Authors treat CSF samples with TransFix for Flow Cytometry analysis within 24 hours. Samples are stained using the EuroFlow small sample tube (SST: CD20, CD45, CD8-surface membrane immunoglobulin (SmIg)lambda, CD56-SmIgkappa, CD4, CD19, SmCD3-CD14, CD38)

http://www.bloodjournal.org/content/123/12/1864.full

Detection & outcome of occult Leptomeningeal disease in diffuse large B-cell lymphoma and Burkitt lymphoma

Sample Type: CSF (diffuse large B-cell lymphoma and Burkitt lymphoma)

Analysis Method: Flow Cytometry

Authors

WH Wilson, JEC Bromberg, M Stetler-Stevenson

Publication

Haematologica. 2014 Jul;99(7):1228-35

Cytology of cerebrospinal fluid (CSF), the diagnostic gold standard has a low sensitivity with reported false negative rate of 20-60% suggesting that pre-treatment involvement of leptomeningeal disease is greater than initially reported. CSF was either analysed within an hour of lumbar puncture, or it was stabilised by two methods, one of which was using TransFix using the method outlined in the paper. Results provided the first evidence that occult CSF involvement by DLBCL or BL is clinically meaningful and associated with an adverse outcome. Results also indicated that patients at risk of CNS disease will benefit from intrathecal chemotherapy.

http://www.ncbi.nlm.nih.gov/pubmed/24727817

Differences in cerebrospinal fluid inflammatory cell reaction of patients with leptomeningeal involvement by Lymphoma and carcinoma

Sample Type: CSF (epithelial cell neoplasia and lymphomas)

Analysis Method: Flow Cytometry

Authors

Julia Illán, Marta Simo, Cristina Serrano, Susana Castañón, Raquel Gonzalo, María Martínez-García, Javier Pardo, Lidia Gómez, Miguel Navarro, Javier Pérez Altozano, Ruth Álvarez, Jordi Bruna, Dolores Subiráemail

Publication

Translational Research ,Volume 164, Issue 6, Pages 460–467

Dissemination of neoplastic cells into the cerebrospinal fluid (CSF) and leptomeninges is a devastating complication in patients with epithelial cell neoplasia and lymphomas. In this study, flow cytometry immunophenotyping was used to analyse CSF leukocyte populations in the CSF of patients diagnosed with leptomeningeal infiltration by solid tumors and lymphomas. CSF samples were collected in tubes containing ethylenediamine tetraacetic acid and an immunofixative reagent (TransFix) for safe transportation and were processed with a median of 3 days from extraction.

http://www.translationalres.com/article/S1931-5244(14)00122-4/pdf

KEY PAPER: Guidelines on the use of multicolor flow cytometry in the diagnosis of haematological neoplasms

Analysis Method: Flow Cytometry

Authors

Ulrika Johansson, David Bloxham, Stephen Couzens, Jennifer Jesson, Ricardo Morilla, Wendy Erber, Marion Macey and British Committee for Standards in Haematology

Publication

British Journal of Haematology, Volume 165, Issue 4, pages 455–488, May 2014

These guidelines have been developed by UK-based clinical flow cytometry (FC) experts along with clinical representatives to take into account advances in the state of the art of clinical flow cytometry since 2002. Their work was revised by the Haemato-oncology Task Force of the British Committee for Standards in Haematology (BCSH) before review by other UK-based Haematologists and FC experts, the BCSH and the British Society for Haematology committee. This document covers a varied range of topics, from selection and setup of flow cytometers through data analysis to training considerations for staff working in the clinical FC laboratory.The guidelines strongly recommend sample age and quality is assessed prior to analysis and in the case of cerebrospinal fluid (CSF), it is recommended that unless immediate analysis is possible then samples can be stored in TransFix/EDTA CSF Sample Storage Tubes for up to 72 hours before analysis*. *TransFix/EDTA CSF Sample Storage Tubes are currently a Research Use Only product and Cytomark recommends that internal validation of the product must be completed by the end user before considering use of the products in a clinical setting.

http://onlinelibrary.wiley.com/doi/10.1111/bjh.12789/abstract

KEY PAPER: Use of TransFix cerebrospinal fluid storage tubes prevents cellular loss and enhances flow cytometric detection of malignant hematological cells after 18 hours of storage

Sample Type: CSF (LHM)

Analysis Method: Flow Cytometry

Authors

A. H. de Jongste, J. Kraan, P. D. van den Broek, R. A. Brooimans, J. E. Bromberg, K. A. van Montfort, P. A. Sillevis Smitt, J. W. Gratama

Publication

Cytometry Part B: Clinical Cytometry Volume 86, Issue 4, pages 272–279, July 2014

In patients with suspected or proven leptomeningeal localised haematological malignancies (LHM) in CSF, rapid assessment is required as cell numbers decline quickly after lumbar puncture. The study aim was to examine leukocyte numbers and detection of LHM by flow cytometry in 99 CSF samples from patients with suspected or proven LHM after 30 minutes and 18 hours of storage in i) TransFix/EDTA CSF Sample Storage Tubes ii) serum-containing medium and iii) no stabilising agent (untreated). It was found that leukocyte numbers in TransFix stabilised CSF were higher than in the corresponding untreated samples at both time points. After 18 hours of storage at 4 degrees Celsius, the detection of LHM was enhanced in TransFix stabilised CSF. For all discordant paired observations, the level of suspicion of a patient having LHM was higher for the TransFix stabilised samples than for the untreated samples. The authors concluded that the use of TransFix/EDTA CSF Sample Storage Tubes prevents cellular loss and enhances flow cytometric detection of LHM after 18 hours of storage.

http://onlinelibrary.wiley.com/doi/10.1002/cyto.b.21097/full

Bone Marrow Stabilisation

KEY PAPER: The impact of stem cells on electron fluxes, proton translation, and ATP synthesis in kidney mitochondria after ischemia/reperfusion

Sample Type: Bone marrow-derived stem cells

Analysis Method: Flow Cytometry

Authors

Beiral HJ1, Rodrigues-Ferreira C, Fernandes AM, Gonsalez SR, Mortari NC, Takiya CM, Sorenson MM, Figueiredo-Freitas C, Galina A, Vieyra A.

Publication

Cell Transplant. 2014 Feb;23(2):207-20

This study aims to show the administration of bone marrow-derived stem cells (BMSCs) can aid recovery of mitochondrial respiration following ischemia/reperfusion (I/R) injuries. The BMSCs were used immediately following isolation from the extracted bone marrow and some were retained for characterisation. 3x106 of the BMSCs were resuspended in PBS and stabilised using 1 part TransFix to 5 parts cell suspension for subsequent immunophenotyping, allowing the researchers to complete other time sensitive aspects of the study.

http://www.ncbi.nlm.nih.gov/pubmed/23211430


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