Highly efficient assay of circulating tumor cells by selective sedimentation with a density gradient medium and microfiltration from whole blood

Jong Myeon Park, June Young Lee, Jeong Gun Lee, Hyoyoung Jeong, Jin Mi Oh, Yeon Jeong Kim, Donghyun Park, Minseok S. Kim, Hun Joo Lee, Jin Ho Oh, Soo Suk Lee, Wonyong Lee, Nam Huh

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

Isolation of circulating tumor cells (CTCs) by size exclusion can yield poor purity and low recovery rates, due to large variations in size of CTCs, which may overlap with leukocytes and render size-based filtration methods unreliable. This report presents a very sensitive, selective, fast, and novel method for isolation and detection of CTCs. Our assay platform consists of three steps: (i) capturing CTCs with anti-EpCAM conjugated microbeads, (ii) removal of unwanted hematologic cells (e.g., leukocytes, erythrocytes, etc.) by selective sedimentation of CTCs within a density gradient medium, and (iii) simple microfiltration to collect these cells. To demonstrate the efficacy of this assay, MCF-7 breast cancer cells (average diameter, 24 μm) and DMS-79 small cell lung cancer cells (average diameter, 10 μm) were used to model CTCs. We investigated the relative sedimentation rates for various cells and/or particles, such as CTCs conjugated with different types of microbeads, leukocytes, and erythrocytes, in order to maximize differences in the physical properties. We observed that greater than 99% of leukocytes in whole blood were effectively removed at an optimal centrifugal force, due to differences in their sedimentation rates, yielding a much purer sample compared to other filter-based methods. We also investigated not only the effect of filtration conditions on recovery rates and sample purity but also the sensitivity of our assay platform. Our results showed a near perfect recovery rate (∼99%) for MCF-7 cells and very high recovery rate (∼89%) for DMS-79 cells, with minimal amounts of leukocytes present.

Original languageEnglish
Pages (from-to)7400-7407
Number of pages8
JournalAnalytical Chemistry
Volume84
Issue number17
DOIs
Publication statusPublished - 2012 Sep 4

Fingerprint

Microfiltration
Sedimentation
Tumors
Assays
Blood
Cells
Recovery
Physical properties

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

Cite this

Park, Jong Myeon ; Lee, June Young ; Lee, Jeong Gun ; Jeong, Hyoyoung ; Oh, Jin Mi ; Kim, Yeon Jeong ; Park, Donghyun ; Kim, Minseok S. ; Lee, Hun Joo ; Oh, Jin Ho ; Lee, Soo Suk ; Lee, Wonyong ; Huh, Nam. / Highly efficient assay of circulating tumor cells by selective sedimentation with a density gradient medium and microfiltration from whole blood. In: Analytical Chemistry. 2012 ; Vol. 84, No. 17. pp. 7400-7407.
@article{2c6085e31f774439885b28a14fc6ac86,
title = "Highly efficient assay of circulating tumor cells by selective sedimentation with a density gradient medium and microfiltration from whole blood",
abstract = "Isolation of circulating tumor cells (CTCs) by size exclusion can yield poor purity and low recovery rates, due to large variations in size of CTCs, which may overlap with leukocytes and render size-based filtration methods unreliable. This report presents a very sensitive, selective, fast, and novel method for isolation and detection of CTCs. Our assay platform consists of three steps: (i) capturing CTCs with anti-EpCAM conjugated microbeads, (ii) removal of unwanted hematologic cells (e.g., leukocytes, erythrocytes, etc.) by selective sedimentation of CTCs within a density gradient medium, and (iii) simple microfiltration to collect these cells. To demonstrate the efficacy of this assay, MCF-7 breast cancer cells (average diameter, 24 μm) and DMS-79 small cell lung cancer cells (average diameter, 10 μm) were used to model CTCs. We investigated the relative sedimentation rates for various cells and/or particles, such as CTCs conjugated with different types of microbeads, leukocytes, and erythrocytes, in order to maximize differences in the physical properties. We observed that greater than 99{\%} of leukocytes in whole blood were effectively removed at an optimal centrifugal force, due to differences in their sedimentation rates, yielding a much purer sample compared to other filter-based methods. We also investigated not only the effect of filtration conditions on recovery rates and sample purity but also the sensitivity of our assay platform. Our results showed a near perfect recovery rate (∼99{\%}) for MCF-7 cells and very high recovery rate (∼89{\%}) for DMS-79 cells, with minimal amounts of leukocytes present.",
author = "Park, {Jong Myeon} and Lee, {June Young} and Lee, {Jeong Gun} and Hyoyoung Jeong and Oh, {Jin Mi} and Kim, {Yeon Jeong} and Donghyun Park and Kim, {Minseok S.} and Lee, {Hun Joo} and Oh, {Jin Ho} and Lee, {Soo Suk} and Wonyong Lee and Nam Huh",
year = "2012",
month = "9",
day = "4",
doi = "10.1021/ac3011704",
language = "English",
volume = "84",
pages = "7400--7407",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "17",

}

Park, JM, Lee, JY, Lee, JG, Jeong, H, Oh, JM, Kim, YJ, Park, D, Kim, MS, Lee, HJ, Oh, JH, Lee, SS, Lee, W & Huh, N 2012, 'Highly efficient assay of circulating tumor cells by selective sedimentation with a density gradient medium and microfiltration from whole blood', Analytical Chemistry, vol. 84, no. 17, pp. 7400-7407. https://doi.org/10.1021/ac3011704

Highly efficient assay of circulating tumor cells by selective sedimentation with a density gradient medium and microfiltration from whole blood. / Park, Jong Myeon; Lee, June Young; Lee, Jeong Gun; Jeong, Hyoyoung; Oh, Jin Mi; Kim, Yeon Jeong; Park, Donghyun; Kim, Minseok S.; Lee, Hun Joo; Oh, Jin Ho; Lee, Soo Suk; Lee, Wonyong; Huh, Nam.

In: Analytical Chemistry, Vol. 84, No. 17, 04.09.2012, p. 7400-7407.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Highly efficient assay of circulating tumor cells by selective sedimentation with a density gradient medium and microfiltration from whole blood

AU - Park, Jong Myeon

AU - Lee, June Young

AU - Lee, Jeong Gun

AU - Jeong, Hyoyoung

AU - Oh, Jin Mi

AU - Kim, Yeon Jeong

AU - Park, Donghyun

AU - Kim, Minseok S.

AU - Lee, Hun Joo

AU - Oh, Jin Ho

AU - Lee, Soo Suk

AU - Lee, Wonyong

AU - Huh, Nam

PY - 2012/9/4

Y1 - 2012/9/4

N2 - Isolation of circulating tumor cells (CTCs) by size exclusion can yield poor purity and low recovery rates, due to large variations in size of CTCs, which may overlap with leukocytes and render size-based filtration methods unreliable. This report presents a very sensitive, selective, fast, and novel method for isolation and detection of CTCs. Our assay platform consists of three steps: (i) capturing CTCs with anti-EpCAM conjugated microbeads, (ii) removal of unwanted hematologic cells (e.g., leukocytes, erythrocytes, etc.) by selective sedimentation of CTCs within a density gradient medium, and (iii) simple microfiltration to collect these cells. To demonstrate the efficacy of this assay, MCF-7 breast cancer cells (average diameter, 24 μm) and DMS-79 small cell lung cancer cells (average diameter, 10 μm) were used to model CTCs. We investigated the relative sedimentation rates for various cells and/or particles, such as CTCs conjugated with different types of microbeads, leukocytes, and erythrocytes, in order to maximize differences in the physical properties. We observed that greater than 99% of leukocytes in whole blood were effectively removed at an optimal centrifugal force, due to differences in their sedimentation rates, yielding a much purer sample compared to other filter-based methods. We also investigated not only the effect of filtration conditions on recovery rates and sample purity but also the sensitivity of our assay platform. Our results showed a near perfect recovery rate (∼99%) for MCF-7 cells and very high recovery rate (∼89%) for DMS-79 cells, with minimal amounts of leukocytes present.

AB - Isolation of circulating tumor cells (CTCs) by size exclusion can yield poor purity and low recovery rates, due to large variations in size of CTCs, which may overlap with leukocytes and render size-based filtration methods unreliable. This report presents a very sensitive, selective, fast, and novel method for isolation and detection of CTCs. Our assay platform consists of three steps: (i) capturing CTCs with anti-EpCAM conjugated microbeads, (ii) removal of unwanted hematologic cells (e.g., leukocytes, erythrocytes, etc.) by selective sedimentation of CTCs within a density gradient medium, and (iii) simple microfiltration to collect these cells. To demonstrate the efficacy of this assay, MCF-7 breast cancer cells (average diameter, 24 μm) and DMS-79 small cell lung cancer cells (average diameter, 10 μm) were used to model CTCs. We investigated the relative sedimentation rates for various cells and/or particles, such as CTCs conjugated with different types of microbeads, leukocytes, and erythrocytes, in order to maximize differences in the physical properties. We observed that greater than 99% of leukocytes in whole blood were effectively removed at an optimal centrifugal force, due to differences in their sedimentation rates, yielding a much purer sample compared to other filter-based methods. We also investigated not only the effect of filtration conditions on recovery rates and sample purity but also the sensitivity of our assay platform. Our results showed a near perfect recovery rate (∼99%) for MCF-7 cells and very high recovery rate (∼89%) for DMS-79 cells, with minimal amounts of leukocytes present.

UR - http://www.scopus.com/inward/record.url?scp=84865734178&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84865734178&partnerID=8YFLogxK

U2 - 10.1021/ac3011704

DO - 10.1021/ac3011704

M3 - Article

C2 - 22881997

AN - SCOPUS:84865734178

VL - 84

SP - 7400

EP - 7407

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 17

ER -