AIM: To investigate the feasibility of separation and cultivation of circulating tumor cells (CTCs) in pancreatic cancer (PaC) using a filtration device. METHODS: In total, 24 PaC patients who were candidates for surgical treatment were enrolled into the study. Peripheral blood samples were collected before an indicated surgery. For each patient, approximately 8 mL of venous blood was drawn from the antecubital veins. A new size-based separation MetaCell technology was used for enrichment and cultivation of CTCs in vitro. (Separated CTCs were cultured on a membrane in FBS enriched RPMI media and observed by inverted microscope. The cultured cells were analyzed by means of histochemistry and immunohistochemistry using the specific antibodies to identify the cell origin. RESULTS: CTCs were detected in 16 patients (66.7%) of the 24 evaluable patients. The CTC positivity did not reflect the disease stage, tumor size, or lymph node involvement. The same percentage of CTC positivity was observed in the metastatic and non-metastatic patients (66.7% vs 66.7%). We report a successful isolation of CTCs in PaC patients capturing proliferating cells. The cells were captured by a capillary action driven size-based filtration approach that enabled cells cultures from the viable CTCs to be unaffected by any antibodies or lysing solutions. The captured cancer cells displayed plasticity which enabled some cells to invade the separating membrane. Further, the cancer cells in the "bottom fraction", may represent a more invasive CTC-fraction. The CTCs were cultured in vitro for further downstream applications. CONCLUSION: The presented size-based filtration method enables culture of CTCs in vitro for possible downstream applications.
- MeSH
- Phenotype MeSH
- Filtration instrumentation MeSH
- Neoplasm Invasiveness MeSH
- Humans MeSH
- Membranes, Artificial * MeSH
- Biomarkers, Tumor metabolism MeSH
- Tumor Cells, Cultured MeSH
- Neoplastic Cells, Circulating metabolism pathology MeSH
- Pancreatic Neoplasms blood metabolism pathology MeSH
- Cell Movement MeSH
- Polycarboxylate Cement MeSH
- Porosity MeSH
- Cell Proliferation MeSH
- Cell Separation instrumentation MeSH
- Feasibility Studies MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
Microreactor technology is an interdisciplinary field that combines science and engineering. This new concept in production, analysis and research is finding increasing application in many different fields. Benefits of this new technology pose a vital influence on chemical industry, biotechnology, the pharmaceutical industry and medicine, life science, clinical and environmental diagnostic. In the last few years, together with microplant development, a great part of research investigation is focused on integrated micro-systems, the so called micro-total-analysis-systems (μ-TAS) or lab-on-chip (LOC). They are devices that perform sampling, sample preparation, detection and date processing in integrated model. Cell sorting, cell lysis, single cell analysis and non-destructive single cell experiments on just one microreactor, makes the LOC platform possible. Clinical diagnostic devices are also leaning towards completely integrated, multiple sophisticated biochemical analyses (PCR amplification, cell lysis, separation and detection) all on a single platform and in real time. Special attention is also paid to the usage of microdevices in tissue. Tissue engineering is one of the most promising fields that can lead to in vitro tissue and organ reconstruction ready for implantation and microdevices can be used to promote the migration, proliferation and the differentiation of cells in controlled situations.
- MeSH
- Biophysics MeSH
- Biomedical Technology MeSH
- Biomedical Research MeSH
- Equipment Design MeSH
- DNA analysis MeSH
- Financing, Organized MeSH
- Immunoassay methods instrumentation MeSH
- Metabolomics methods MeSH
- Patch-Clamp Techniques instrumentation MeSH
- Microchemistry instrumentation MeSH
- Microchip Analytical Procedures MeSH
- Microfluidics methods instrumentation MeSH
- Microfluidic Analytical Techniques methods MeSH
- Microtechnology methods instrumentation MeSH
- Miniaturization methods instrumentation MeSH
- Polymerase Chain Reaction methods instrumentation MeSH
- Cell Separation methods instrumentation MeSH
- Tissue Engineering methods instrumentation MeSH
- Publication type
- Review MeSH
BACKGROUND: A product with well-preserved haemostatic function of platelets is the ultimate goal of platelet concentrate production. However, platelet activation and apoptosis are induced by both collection and storage of platelet concentrates. AIM OF STUDY: Platelet concentrates obtained either by two blood separators with different technology of apheresis (Haemonetics MCS+, Haemonetics Corp. Braintree, USA and Trima Accel, Gambro BCT Inc., Lakewood, USA, respectively) or derived from buffy-coat were compared using evaluation of pH, LDH, lactate, glucose, annexin V, and sP-selectin levels immediately after collecting and at the end of expiration to estimate the differences in the activation and apoptosis of platelets in these products. RESULTS: The lowest degree of platelet activation was found in products obtained by Haemonetics MCS+ apparatus at the time of collection. Platelet concentrates obtained by apheresis revealed higher rise of LDH, annexin V and sP-selectin compared to buffy-coat derived platelets. Products from Haemonetics MCS+ showed higher rise of annexin V in comparison with products from Trima separator. Increase of LDH and sP-selectin in both apheresis products was comparable. CONCLUSIONS: On the basis of changes of sP-selectin and annexin V levels it could be concluded that initial platelet activation, which is induced by apheresis, is very likely without any further impact on quality of platelets during storage. Development of platelet storage lesions is influenced especially by storage conditions and platelet concentration in products.
- MeSH
- Platelet Activation physiology MeSH
- Apoptosis physiology MeSH
- Biomarkers blood MeSH
- Blood Donors MeSH
- Adult MeSH
- Financing, Organized MeSH
- Blood Preservation MeSH
- Middle Aged MeSH
- Humans MeSH
- Platelet Count MeSH
- Prospective Studies MeSH
- Reference Values MeSH
- Reproducibility of Results MeSH
- Cell Separation methods instrumentation MeSH
- Blood Component Removal methods instrumentation MeSH
- Blood Platelets physiology MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Female MeSH
- Publication type
- Evaluation Study MeSH
- Comparative Study MeSH
