Membrane microfilter device for selective capture, electrolysis and genomic analysis of human circulating tumor cells.

Abstract

This paper presents development of a parylene membrane microfilter device for single stage capture and electrolysis of circulating tumor cells (CTCs) in human blood, and the potential of this device to allow genomic analysis. The presence and number of CTCs in blood has recently been demonstrated to provide significant prognostic information for patients with metastatic breast cancer. While finding as few as five CTCs in about 7.5mL of blood (i.e., 10(10) blood cells in) is clinically significant, detection of CTCs is currently difficult and time consuming. CTC enrichment is performed by either gradient centrifugation of CTC based on their buoyant density or magnetic separation of epithelial CTC, both of which are laborious procedures with variable efficiency, and CTC identification is typically done by trained pathologists through visual observation of stained cytokeratin-positive epithelial CTC. These processes may take hours, if not days. Work presented here provides a micro-electro-mechanical system (MEMS)-based option to make this process simpler, faster, better and cheaper. We exploited the size difference between CTCs and human blood cells to achieve the CTC capture on filter with approximately 90% recovery within 10 min, which is superior to current approaches. Following capture, we facilitated polymerase chain reaction (PCR)-based genomic analysis by performing on-membrane electrolysis with embedded electrodes reaching each of the individual 16,000 filtering pores. The biggest advantage for this on-membrane in situ cell lysis is the high efficiency since cells are immobilized, allowing their direct contact with electrodes. As a proof-of-principle, we show beta actin gene PCR, the same technology can be easily extended to real time PCR for CTC-specific transcript to allow molecular identification of CTC and their further characterization.

• Authors: Siyang Zheng, Henry Lin, Jing-Quan Liu, Marija Balic, Ram Datar, Richard J Cote, Yu-Chong Tai
• Journal: Journal of chromatography. A (J Chromatogr A) Vol. 1162 Issue 2 Pg. 154-61 (Aug 31 2007) ISSN: 0021-9673 [Print] Netherlands
• PMID: 17561026 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Membranes, Artificial
• Polymers
• Xylenes
• parylene
• Keratins

Topics

• Cell Line, Tumor
• Electrolysis (instrumentation, methods)
• Genomics (instrumentation, methods)
• Humans
• Immunohistochemistry
• Keratins (analysis)
• Male
• Membranes, Artificial
• Microscopy, Electron, Scanning
• Neoplastic Cells, Circulating (metabolism, pathology, ultrastructure)
• Polymerase Chain Reaction
• Polymers (chemistry)
• Xylenes (chemistry)