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Two-Step Acoustophoresis Separation of Live Tumor Cells from Whole Blood

Undvall Anand, Eva LU ; Magnusson, Cecilia LU ; Lenshof, Andreas LU ; Ceder, Yvonne LU orcid ; Lilja, Hans LU orcid and Laurell, Thomas LU (2021) In Analytical Chemistry 93(51). p.17076-17085
Abstract

There is an unmet clinical need to extract living circulating tumor cells (CTCs) for functional studies and in vitro expansion to enable drug testing and predict responses to therapy in metastatic cancer. Here, we present a novel two-step acoustophoresis (A2) method for isolation of unfixed, viable cancer cells from red blood cell (RBC) lysed whole blood. The A2 method uses an initial acoustofluidic preseparation step to separate cells based on their acoustic mobility. This acoustofluidic step enriches viable cancer cells in a central outlet, but a significant number of white blood cells (WBCs) remain in the central outlet fraction due to overlapping acoustophysical properties of these viable cells. A subsequent purging step was... (More)

There is an unmet clinical need to extract living circulating tumor cells (CTCs) for functional studies and in vitro expansion to enable drug testing and predict responses to therapy in metastatic cancer. Here, we present a novel two-step acoustophoresis (A2) method for isolation of unfixed, viable cancer cells from red blood cell (RBC) lysed whole blood. The A2 method uses an initial acoustofluidic preseparation step to separate cells based on their acoustic mobility. This acoustofluidic step enriches viable cancer cells in a central outlet, but a significant number of white blood cells (WBCs) remain in the central outlet fraction due to overlapping acoustophysical properties of these viable cells. A subsequent purging step was employed to remove contaminating WBCs through negative selection acoustophoresis with anti-CD45-functionalized negative acoustic contrast particles. We processed 1 mL samples of 1:1 diluted RBC lysed whole blood mixed with 10 000 DU145 cells through the A2 method. Additional experiments were performed using 1000 DU145 cells spiked into 1.5 × 106 WBCs in 1 mL of buffer to further elucidate the dynamic range of the method. Using samples with 10 000 DU145 cells, we obtained 459 ± 188-fold depletion of WBC and 42% recovery of viable cancer cells. Based on spiked samples with 1000 DU145 cells, our cancer cell recovery was 28% with 247 ± 156-fold WBC depletion corresponding to a depletion efficacy of ≥99.5%. The novel A2 method provides extensive elimination of WBCs combined with the gentle recovery of viable cancer cells suitable for downstream functional analyses and in vitro culture.

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author
; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Acoustics, Cell Separation, Humans, Leukocyte Count, Leukocytes, Neoplastic Cells, Circulating
in
Analytical Chemistry
volume
93
issue
51
pages
10 pages
publisher
The American Chemical Society (ACS)
external identifiers
  • pmid:34913344
  • scopus:85121987180
ISSN
1520-6882
DOI
10.1021/acs.analchem.1c04050
language
English
LU publication?
yes
id
40bed163-923c-4457-b232-8ac6afa40036
date added to LUP
2022-01-17 13:06:58
date last changed
2024-06-15 23:59:03
@article{40bed163-923c-4457-b232-8ac6afa40036,
  abstract     = {{<p>There is an unmet clinical need to extract living circulating tumor cells (CTCs) for functional studies and in vitro expansion to enable drug testing and predict responses to therapy in metastatic cancer. Here, we present a novel two-step acoustophoresis (A2) method for isolation of unfixed, viable cancer cells from red blood cell (RBC) lysed whole blood. The A2 method uses an initial acoustofluidic preseparation step to separate cells based on their acoustic mobility. This acoustofluidic step enriches viable cancer cells in a central outlet, but a significant number of white blood cells (WBCs) remain in the central outlet fraction due to overlapping acoustophysical properties of these viable cells. A subsequent purging step was employed to remove contaminating WBCs through negative selection acoustophoresis with anti-CD45-functionalized negative acoustic contrast particles. We processed 1 mL samples of 1:1 diluted RBC lysed whole blood mixed with 10 000 DU145 cells through the A2 method. Additional experiments were performed using 1000 DU145 cells spiked into 1.5 × 106 WBCs in 1 mL of buffer to further elucidate the dynamic range of the method. Using samples with 10 000 DU145 cells, we obtained 459 ± 188-fold depletion of WBC and 42% recovery of viable cancer cells. Based on spiked samples with 1000 DU145 cells, our cancer cell recovery was 28% with 247 ± 156-fold WBC depletion corresponding to a depletion efficacy of ≥99.5%. The novel A2 method provides extensive elimination of WBCs combined with the gentle recovery of viable cancer cells suitable for downstream functional analyses and in vitro culture.</p>}},
  author       = {{Undvall Anand, Eva and Magnusson, Cecilia and Lenshof, Andreas and Ceder, Yvonne and Lilja, Hans and Laurell, Thomas}},
  issn         = {{1520-6882}},
  keywords     = {{Acoustics; Cell Separation; Humans; Leukocyte Count; Leukocytes; Neoplastic Cells, Circulating}},
  language     = {{eng}},
  month        = {{12}},
  number       = {{51}},
  pages        = {{17076--17085}},
  publisher    = {{The American Chemical Society (ACS)}},
  series       = {{Analytical Chemistry}},
  title        = {{Two-Step Acoustophoresis Separation of Live Tumor Cells from Whole Blood}},
  url          = {{http://dx.doi.org/10.1021/acs.analchem.1c04050}},
  doi          = {{10.1021/acs.analchem.1c04050}},
  volume       = {{93}},
  year         = {{2021}},
}