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
- fenotyp MeSH
- filtrace přístrojové vybavení MeSH
- invazivní růst nádoru MeSH
- lidé MeSH
- membrány umělé * MeSH
- nádorové biomarkery metabolismus MeSH
- nádorové buňky kultivované MeSH
- nádorové cirkulující buňky metabolismus patologie MeSH
- nádory slinivky břišní krev metabolismus patologie MeSH
- pohyb buněk MeSH
- polykarboxylátový cement MeSH
- poréznost MeSH
- proliferace buněk MeSH
- separace buněk přístrojové vybavení MeSH
- studie proveditelnosti MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem 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
- biofyzika MeSH
- biomedicínské technologie MeSH
- biomedicínský výzkum MeSH
- design vybavení MeSH
- DNA analýza MeSH
- financování organizované MeSH
- imunoanalýza metody přístrojové vybavení MeSH
- metabolomika metody MeSH
- metoda terčíkového zámku přístrojové vybavení MeSH
- mikrochemie přístrojové vybavení MeSH
- mikročipové analytické postupy MeSH
- mikrofluidika metody přístrojové vybavení MeSH
- mikrofluidní analytické techniky metody MeSH
- mikrotechnologie metody přístrojové vybavení MeSH
- miniaturizace metody přístrojové vybavení MeSH
- polymerázová řetězová reakce metody přístrojové vybavení MeSH
- separace buněk metody přístrojové vybavení MeSH
- tkáňové inženýrství metody přístrojové vybavení MeSH
- Publikační typ
- přehledy 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
- aktivace trombocytů fyziologie MeSH
- apoptóza fyziologie MeSH
- biologické markery krev MeSH
- dárci krve MeSH
- dospělí MeSH
- financování organizované MeSH
- konzervace krve MeSH
- lidé středního věku MeSH
- lidé MeSH
- počet trombocytů MeSH
- prospektivní studie MeSH
- referenční hodnoty MeSH
- reprodukovatelnost výsledků MeSH
- separace buněk metody přístrojové vybavení MeSH
- separace krevních složek metody přístrojové vybavení MeSH
- trombocyty fyziologie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- hodnotící studie MeSH
- srovnávací studie MeSH
