Analytical Chemistry, 2014: Single-Cell Copy Number Analysis of Prostate Cancer Cells Captured with Geometrically Enhanced Differential Immunocapture Microdevices
Citation: Pratt ED, Stepansky A, Hicks J, Kirby BJ. Single-Cell Copy Number Analysis of Prostate Cancer Cells Captured with Geometrically Enhanced Differential Immunocapture Microdevices, Gastroenterology 2014;146:647-651. doi pdf
Abstract: Limited access to tumor tissue makes repeated sampling and real-time tracking of cancer progression infeasible. Circulating tumor cells (CTCs) provide the capacity for real-time genetic characterization of a disseminating tumor cell population via a simple blood draw. However, there is no straightforward method to analyze broadscale genetic rearrangements in this heterogeneous cell population at the single cell level. We present a one-step controllable chemical extraction of whole nuclei from prostate cancer cells captured using geometrically enhanced differential immunocapture (GEDI) microdevices. We have successfully used copy number profile analysis to differentiate between two unique cancer cell line populations of metastatic origin (LNCaP and VCaP) and to analyze key mutations important in disease progression.
Figures:
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- Figure 1. (A) LNCaP nuclei were sorted via three methods: (1) FACS only, (2) capture in GEDI microdevices, followed by serial dilution, and (3) serial dilution only. VCaP controls underwent FACS. (B) Onchip lysis of LNCaP cells in a GEDI microdevice via NST-DAPI buffer, prior to elution via flow. Digestion of labeled cell membrane (green) and simultaneous nucleus labeling (red) is easily observed. Scale bars are 100 micron. doi pdf
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- Figure 2. (A) Copy-number profiles of individual LNCaP and VCaP nuclei after undergoing amplification followed by single-nucleus sequencing. (B) By comparing copy-number profiles of the two different prostate cancer cell lines, one can contrast unique genetic amplifications and deletions across their respective genomes. CNVs of interest are marked by a solid gray line. doi pdf
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- Figure 3. (A) Simple linear regression was used to determine r2 values for copy-number profile comparisons. (B) Dendrogram showing level of genetic relationship between cell populations. The (2) indicates profiles averaged over two nuclei, all others are from single cells. doi pdf