High‐Purity Prostate Circulating Tumor Cell Isolation by a Polymer Nanofiber‐Embedded Microchip for Whole Exome Sequencing

Zhao, Libo; Lu, Yi‐Tsung; Li, Fuqiang; Wu, Kui; Hou, Shuang; Yu, Juehua; Shen, Qinglin; Wu, Danni; Song, Min; OuYang, Wei‐Han; Luo, Zheng; Lee, Tom; Fang, Xiaohong; Shao, Chen; Xu, Xun; Garcia, Mitch A.; Chung, Leland W.K.; Rettig, Matthew B.; Tseng, Hsian‐Rong; Posadas, Edwin M.

doi:10.1002/adma.201205237

Cited by 144 publications

(151 citation statements)

References 24 publications

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“…13,14 Briefly, poly(lactic-co-glycolic acid) nano-spun chips, manufactured by an electro-spin method in our nano-materials laboratory, were assembled onto a laser microdissection slide (Leica Microsystems Inc.) with an overlaid polydimethylsiloxane microfluidic component and attached to a syringe-based microfluidic pump (KD Scientific, Holliston, MA). Prepared samples were injected through the device at the previously established optimal flow rate of 0.5 mL/hour and were then fixed using 100% ethanol.…”

Section: Nanovelcro/lcm Ctc Chip Processingmentioning

confidence: 99%

Reality of Single Circulating Tumor Cell Sequencing for Molecular Diagnostics in Pancreatic Cancer

Court

Ankeny

Sho

et al. 2016

The Journal of Molecular Diagnostics

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To understand the potential and limitations of circulating tumor cell (CTC) sequencing for molecular diagnostics, we investigated the feasibility of identifying the ubiquitous KRAS mutation in single CTCs from pancreatic cancer (PC) patients. We used the NanoVelcro/laser capture microdissection CTC platform, combined with whole genome amplification and KRAS Sanger sequencing. We assessed both KRAS codon-12 coverage and the degree that allele dropout during whole genome amplification affected the detection of KRAS mutations from single CTCs. We isolated 385 single cells, 163 from PC cell lines and 222 from the blood of 12 PC patients, and obtained KRAS sequence coverage in 218 of 385 single cells (56.6%). For PC cell lines with known KRAS mutations, single mutations were detected in 67% of homozygous cells but only 37.4% of heterozygous single cells, demonstrating that both coverage and allele dropout are important causes of mutation detection failure from single cells. We could detect KRAS mutations in CTCs from 11 of 12 patients (92%) and 33 of 119 single CTCs sequenced, resulting in a KRAS mutation detection rate of 27.7%. Importantly, KRAS mutations were never found in the 103 white blood cells sequenced. Sequencing of groups of cells containing between 1 and 100 cells determined that at least 10 CTCs are likely required to reliably assess KRAS mutation status from CTCs. (J Mol Diagn 2016, 18: 688e696; http://dx

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Section: Nanovelcro/lcm Ctc Chip Processingmentioning

confidence: 99%

Reality of Single Circulating Tumor Cell Sequencing for Molecular Diagnostics in Pancreatic Cancer

Court

Ankeny

Sho

et al. 2016

The Journal of Molecular Diagnostics

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“…For instance, when the cell analysis of a tumor bulk demands a high accuracy and sensitivity or in the cases where differentiation of viable and non-viable cells is required, using a microchip is almost inevitable (Zhao et al, 2013). The microchips which currently used specifically for single cell isolation generally work based on three main principles: Droplet-in-oil-based isolation (Brouzes et al, 2009), Hydrodynamic cell traps (Di Carlo et al, 2006), and Pneumatic membrane valving (GomezSjoberg et al, 2007).…”

Section: Single Cell Isolationmentioning

confidence: 99%

“…Later in 2013, NCI-1650 antigen was applied as an immunologic target with a nitrocellulose membrane as scaffold to isolate the CTCs of lung cancer . In the same year Zhao et al designed a system called "NanoVelcro Chip" coupled with ArcturusXT™ laser capture microdissection (LCM) technology to isolate the CTCs of prostate cancer for the next generation sequencing [NGS] purposes (Zhao et al, 2013). In 2014, a modern microchip named "geometrically enhanced mixing (GEM) chip" was successfully used to isolate the pancreatic cancer CTCs with a great specificity and accuracy (Sheng et al, 2014).…”

Section: 879 Microchips and Their Significance In Isolation Of Circumentioning

confidence: 99%

Microchips and their Significance in Isolation of Circulating Tumor Cells and Monitoring of Cancers

Sahmani

Vatanmakanian²,

Goudarzi³

et al. 2016

Asian Pacific Journal of Cancer Prevention

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“…[89] Eine Reihe von Untersuchungen befassten sich mit der Sequenzierung von Patienten-CTCs und ergaben, dass charakteristische Mutationen, die im Primärtumor vorlagen, auch in den zirkulierenden Zellen nachgewiesen werden konnten. [64,[90][91][92] Der Inhalt des CTC-Invasoms bleibt jedoch unbekannt. CTCs sind inhärent zerbrechliche Zellen, was ihre Analyse erschwert, weil es schwierig ist zu beurteilen, ob Zellen Veränderungen in ihrem molekularen Profil durchlaufen, wenn sie zwischen den Fängervorrichtungen und den Analysegeräten transferiert werden.…”

Section: Integrierte Molekulare Analyse Von Ctcsunclassified

Profilierung zirkulierender Tumorzellen mit Apparaturen und Materialien der nächsten Generation

Green

Safaei²,

Mepham

et al. 2015

Angewandte Chemie

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In den letzten zehn Jahren gab es wichtige Fortschritte bei der Erfassung seltener zirkulierender Tumorzellen (CTCs) aus dem Blut von Krebspatienten als eine entscheidende Voraussetzung für die nichtinvasive Tumorprofilierung. Diese Fortschritte haben zudem neue Einblicke in die Materialchemie und die Mikrofluidik erbracht und ermöglichten die Erfassung und Auszählung von CTCs mit beispielloser Empfindlichkeit. Allerdings wurde auch immer klarer, dass einfaches Isolieren und Auszählen von Tumorzellen, die in den Kreislauf gelangt waren, nicht die benötigten Informationen lieferte, um unser Verständnis der Biologie dieser seltenen Zellen zu vertiefen oder um sie sich in der Therapie besser zunutze zu machen. Mithilfe von Apparaturen und Materialien der nächsten Generation mit maßgeschneiderten physikalischen und chemischen Eigenschaften ist man inzwischen in der Lage, mehr Informationen aus CTCs zu gewinnen. In diesem Kurzaufsatz werden die Arbeiten der letzten zehn Jahre in diesem Gebiet diskutiert, wobei insbesondere die bahnbrechenden Studien der letzten fünf Jahre Berücksichtigung finden, die den Blick über das einfache Isolieren von CTCs hinaus auf Ansätze gerichtet haben, die eine tiefergehende Analyse ermöglichen.

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High‐Purity Prostate Circulating Tumor Cell Isolation by a Polymer Nanofiber‐Embedded Microchip for Whole Exome Sequencing

Cited by 144 publications

References 24 publications

Reality of Single Circulating Tumor Cell Sequencing for Molecular Diagnostics in Pancreatic Cancer

Reality of Single Circulating Tumor Cell Sequencing for Molecular Diagnostics in Pancreatic Cancer

Microchips and their Significance in Isolation of Circulating Tumor Cells and Monitoring of Cancers

Profilierung zirkulierender Tumorzellen mit Apparaturen und Materialien der nächsten Generation

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