Applications of TransFix: Stabilisation of Bone Marrow for Flow Cytometry
Bone marrow (BM) examination is an essential procedure for the evaluation of a variety of clinical haematologic disorders and for preclinical toxicity studies of anti-cancer compounds and immunosuppressive agents. This has resulted in bone marrow flow cytometry becoming a routine test in clinical laboratories.
Independent research involving the use of TransFix to stabilise bone marrow samples has been published, including the use of TransFix for the enumeration of bone marrow mast cells (case study 1), for immunophenotyping rat bone marrow stem cells (case study 2), and for transporting bone marrow for cancer diagnosis (case study 3).
Benefits of TransFix
In cases in which sample processing can not be conducted within the first 24 hours following a bone marrow puncture, TransFix has been used to avoid deterioration of cells. Stabilising bone marrow stem cells with TransFix prior to immunophenotyping has allowed researchers to complete time sensitive aspects of studies.
Please note that Cytomark have not validated the use of TransFix for bone marrow for IVD use.
Case Study 1: Use of Transfix for the Enumeration of Bone Marrow Mast Cells
Mastocytosis is a term used to designate a heterogeneous group of disorders characterized by an abnormal proliferation and accumulation of mast cells (MCs). Aberrant mast cell surface marker expression can be detected in the bone marrow aspirate by flow cytometry.
For the enumeration of Bone Marrow MCs, fresh samples with high cell viability (>95%) should be used. In cases in which it is not expected to perform sample processing within the first 24 h following a BM puncture, a stabilizing solution should be used to avoid deterioration of cells [e.g., TransFix (Cytomark, Buckingham, UK)].
Case Study 2: Stabilisation of Rat BMSCs with TransFix for Immunophenotyping
This study investigated how 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. 3×106 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.
Reference: Beiral et al. (2014) The Impact of Stem Cells on Electron Fluxes, Proton Translocation, and ATP Synthesis in Kidney Mitochondria After Ischemia/Reperfusion. Cell Transplantation (23): 207-220
Case Study 3: Use of TransFix to Support Transportation of Bone Marrow Samples for Cancer Diagnosis
This poster details how modern transport and telecommunications technology can facilitate cancer programmes in resource-poor areas. TransFix was used to transport bone marrow samples from Tanzania to Dublin, Ireland (2 day transit time) where interim diagnostic services were provided while helping to develop local paediatric cancer diagnostic capacity. Marrow samples collected in TransFix showed some loss of cytoplasmic antigens but sufficiently preserved membrane antigens to permit interpretation.
Reference: McDermott et al. (2016) The utilisation of modern transport and telecommunications platforms to assist in the remote provision of pediatric cancer diagnostics in Tanzania. POSTER presented at International Society of Paediatric Oncology.
- BM is taken into EDTA
- Perform WBC count on BM sample (beware of clots!)
- Dilute in PBS to ensure WBC < 40 109/L
- Label TRANSFIX tube with above patient details
- Add 1.5 ml BM to labelled transfix tube – mix gently by inversion X 5
- Repeat for 2 more tubes if sufficient BM available
- Send via DHL as soon as possible
TransFix is available in 1ml or 20ml vials, or in TransFix Sample Storage Tubes which contain 0.2ml TransFix for the addition of 1ml of sample. If you have any questions regarding the use of TransFix for bone marrow, please contact us.