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Sensitive cytometry based system for enumeration, capture and analysis of gene mutations of circulating tumor cells.

Sawada T, Watanabe M, Fujimura Y, Yagishita S, Shimoyama T, Maeda Y, Kanda S, Yunokawa M, Tamura K, Tamura T, Minami H, Koh Y, Koizumi F - Cancer Sci. (2016)

Bottom Line: We compared the number of CTC detected using our system with those detected using the CellSearch system in 46 patients with various cancers.The percentage of samples with detected CTC was significantly higher with our system (65.2%) than with CellSearch (28.3%).The number of detected CTC per patient using our system was statistically higher than that using CellSearch (median 5, 0; P = 0.000172, Wilcoxon test).

View Article: PubMed Central - PubMed

Affiliation: Shien-Lab, National Cancer Center Hospital, Tokyo, Japan.

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Related in: MedlinePlus

Circulating tumor cell (CTC) enrichment process of our system. Peripheral blood is collected in 5 mL EDTA test tube. 4 mL of blood is hemolyzed using BD Pharm Lyse buffer. Subsequently, Dynabeads CD45 eliminates leukocytes from the samples. Processed samples are fixed with BD Phosflow and stained with FITC‐conjugated anti‐CK, PE‐conjugated anti‐EpCAM and Alexa Fluor 700‐conjugated anti‐CD45 antibodies. After incubation overnight, samples are applied to FISHMAN‐R or On‐chip Sort.
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cas12868-fig-0002: Circulating tumor cell (CTC) enrichment process of our system. Peripheral blood is collected in 5 mL EDTA test tube. 4 mL of blood is hemolyzed using BD Pharm Lyse buffer. Subsequently, Dynabeads CD45 eliminates leukocytes from the samples. Processed samples are fixed with BD Phosflow and stained with FITC‐conjugated anti‐CK, PE‐conjugated anti‐EpCAM and Alexa Fluor 700‐conjugated anti‐CD45 antibodies. After incubation overnight, samples are applied to FISHMAN‐R or On‐chip Sort.

Mentions: Circulating tumor cell enrichment from 4 mL of peripheral blood includes hemolysis and elimination of leukocytes by using CD45 antibody‐conjugated microbeads. After the enrichment process, the CTC‐enriched samples were subsequently fixed and stained using anti‐CK‐FITC, anti‐EpCAM‐PE and anti‐CD45‐Alexa Fluor 700 antibodies. Then, the samples were applied to FISHMAN‐R or On‐chip Sort. A flowchart of this process including the CTC enrichment percentage is shown in Figure 2.


Sensitive cytometry based system for enumeration, capture and analysis of gene mutations of circulating tumor cells.

Sawada T, Watanabe M, Fujimura Y, Yagishita S, Shimoyama T, Maeda Y, Kanda S, Yunokawa M, Tamura K, Tamura T, Minami H, Koh Y, Koizumi F - Cancer Sci. (2016)

Circulating tumor cell (CTC) enrichment process of our system. Peripheral blood is collected in 5 mL EDTA test tube. 4 mL of blood is hemolyzed using BD Pharm Lyse buffer. Subsequently, Dynabeads CD45 eliminates leukocytes from the samples. Processed samples are fixed with BD Phosflow and stained with FITC‐conjugated anti‐CK, PE‐conjugated anti‐EpCAM and Alexa Fluor 700‐conjugated anti‐CD45 antibodies. After incubation overnight, samples are applied to FISHMAN‐R or On‐chip Sort.
© Copyright Policy - creativeCommonsBy-nc-nd
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4814266&req=5

cas12868-fig-0002: Circulating tumor cell (CTC) enrichment process of our system. Peripheral blood is collected in 5 mL EDTA test tube. 4 mL of blood is hemolyzed using BD Pharm Lyse buffer. Subsequently, Dynabeads CD45 eliminates leukocytes from the samples. Processed samples are fixed with BD Phosflow and stained with FITC‐conjugated anti‐CK, PE‐conjugated anti‐EpCAM and Alexa Fluor 700‐conjugated anti‐CD45 antibodies. After incubation overnight, samples are applied to FISHMAN‐R or On‐chip Sort.
Mentions: Circulating tumor cell enrichment from 4 mL of peripheral blood includes hemolysis and elimination of leukocytes by using CD45 antibody‐conjugated microbeads. After the enrichment process, the CTC‐enriched samples were subsequently fixed and stained using anti‐CK‐FITC, anti‐EpCAM‐PE and anti‐CD45‐Alexa Fluor 700 antibodies. Then, the samples were applied to FISHMAN‐R or On‐chip Sort. A flowchart of this process including the CTC enrichment percentage is shown in Figure 2.

Bottom Line: We compared the number of CTC detected using our system with those detected using the CellSearch system in 46 patients with various cancers.The percentage of samples with detected CTC was significantly higher with our system (65.2%) than with CellSearch (28.3%).The number of detected CTC per patient using our system was statistically higher than that using CellSearch (median 5, 0; P = 0.000172, Wilcoxon test).

View Article: PubMed Central - PubMed

Affiliation: Shien-Lab, National Cancer Center Hospital, Tokyo, Japan.

Show MeSH
Related in: MedlinePlus