Vita-Cap™ C1 Kit contains essential components to capture CTCs, iCTCs, CECs and hematopoietic immune cells in up to 3-mL heparinized blood, and to prepare for downstream analyses of iCTCs including culture, flow cytometry phenotyping, gene expression profiling and NGS.
Vita-Cap™ C1: Processes 2-ml of whole blood
To capture rare cells from whole blood, venous blood is collected in a standard blood collection heparin tube. The specimen is either transported immediately or stored/shipped at 2-8°C within 48 hours to the laboratory for processing. Optimally, 2-mL of blood is introduced into the Vita-Cap™ tube that contained 6-mL anticoagulant cell dilute working buffer. The sealed tube is then rotated on a tube roller at slow rotation speed, 5rpm, for 4 hours. Unlike the majority of blood cells, tumor, endothelial and immune progenitor cells, particularly those cells which are tumorous, epithelial, endothelial and stem-like in origin, differentially attach themselves to the CAM-coated beads (scaffolds) on the inner surface of the Vita-Cap™ tube. The majority of blood cells remains floating and is subsequently washed off of the tube inner surface. Rare cells, such as CTCs, hematopoietic immune cells, and circulating endothelial cells (CECs) are thus concentrated 1 million-fold from original whole blood.
Once the tube is washed, DNA and RNA of the attached CAM-avid tumor, endothelial and immune progenitor cells can be extracted for proteomics, gene expression profiling and genomic analyses. Alternatively, after washing of the tube, CAM-avid cells can be released from CAM scaffolds by enzymatic digestion, and the identity of the enriched cells characterized by multi-parameter flow cytometry or fluorescent microscopy. Using multiplex flow cytometry protocol with EPI+, TP+, 7AAD+ and HL- (hematopoietic lineage), enumeration of CTCs can be performed.
Workflow for Blood Collection, CAM Cell Enrichment and Flow Cytometry Enumeration of CTCs in Minimum Volume of a Patient’s Blood.
Recent studies show that proliferative and invasion-competent circulating tumor cells (iCTCs) are invaluable in many downstream applications that depend on the ability of the isolated tumor cells to recapitulate tumor growth and metastasis in ex vivo culture and xenograft mouse models. Vita-Cap™ C1 tube is intended to capture, identify, enumerate and culture CTCs and hematopoietic (endothelial) progenitor cells in cancer patient’s blood. Cells captured in Vita-Cap™ C1 tube can be cultured in the same tube up to 28 days, allowing significant amplification of iCTCs and reduction of WBCs for xenograft mouse models and various downstream analyses.
Workflow for Blood Collection, CAM Cell Enrichment and Continued Culture of CTCs in Minimum Volume of a Patient’s Blood.
Workflow for Blood Collection, CAM Cell Enrichment and Molecular Analyses of CTCs in Minimum Volume of a Patient’s Blood.