Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Sep 7;54(37):10838-42.
doi: 10.1002/anie.201505294. Epub 2015 Jul 15.

High-Throughput, Label-Free Isolation of Cancer Stem Cells on the Basis of Cell Adhesion Capacity

Yuanqing Zhang  1   2 , Minhao Wu  3 , Xin Han  1   2 , Ping Wang  4 , Lidong Qin  5   6
Affiliations

High-Throughput, Label-Free Isolation of Cancer Stem Cells on the Basis of Cell Adhesion Capacity

Yuanqing Zhang et al. Angew Chem Int Ed Engl. .

Abstract

Herein we report a microfluidics method that enriches cancer stem cells (CSCs) or tumor-initiating cells on the basis of cell adhesion properties. In our on-chip enrichment system, cancer cells were driven by hydrodynamic forces to flow through microchannels coated with basement membrane extract. Highly adhesive cells were captured by the functionalized microchannels, and less adhesive cells were collected from the outlets. Two heterogeneous breast cancer cell lines (SUM-149 and SUM-159) were successfully separated into enriched subpopulations according to their adhesive capacity, and the enrichment of the cancer stem cells was confirmed by flow cytometry biomarker analysis and tumor-formation assays. Our findings show that the less adhesive phenotype is associated with a higher percentage of CSCs, higher cancer-cell motility, and higher resistance to chemotherapeutic drugs.

Keywords: biomaterials; cancer stem cells; cell adhesion; high-throughput methods; microfluidics.

PubMed Disclaimer

Figures

Fig. 1
Fig. 1
(A) Scheme (upper) and photograph (lower) of the high-throughput cell adhesion chip (HCA-Chip). The chip was filled with red dye. (B) Optical microscopic images of the HCA-Chip. The depth of the microchannel is approximately 25 μm. Scale bar: 30 μm. (C) Optical microscopic images of the microchannel’s structure and captured MCF-7 cell. Scale bar: 20 μm.
Fig. 2
Fig. 2
Characterization of the HCA-Chip cell capture yield with cancer cells in buffer solution. (A) Dependence of MCF-7 capture efficiency on flow rate. (B) Capture of MCF-7 cells in HCA-Chips that were coated with different biomaterials. (C) Comparison of the capture yield of different cell lines in the BME-coated HCA-Chip at different flow rates. Error bars represent the standard deviation of three replicates.
Fig. 3
Fig. 3
Analyses of the subpopulations of separated cells. (A) Mammospheres derived from SUM-149 cells of low and high adhesion. (B) Mammosphere formation efficiency of subpopulations of low and high adhesion. All three replicates showed consistently greater efficiency in flexible cells. (C) Scatter plot of CD44/CD24 expression in SUM-149 cells before and after HCA-Chip separation. (D) Heat map showing the expression levels of genes related to cell-substrate adhesion, chemotaxis, and cell motion in cells of low and high adhesion. (E) Western blot analysis showing E-cadherin and Vinmentin production. (F) Low-adhesive cells have higher cell invasive capacity. (G) Resistance of lower and higher adhesive cells to different concentrations of doxorubicin (DOX). The subpopulations of cells were separated 3 times by different HCA-Chips.
Fig. 4
Fig. 4
Low-adhesive B16 melanoma cells demonstrate increased potential to induce tumor development. (A) Representative images of lungs are shown. (B) Numbers of tumor foci and total cells in the lung, as well as lung weights, were examined in B6 mice that were challenged with high-, medium-, or low-adhesive B16 cells. (C) Tissue pathology in the lungs of B6 mice was assessed by hematoxylin and eosin staining after challenge with low-, medium-, or high-adhesive B16 cells. Magnification, ×400. (D) Expression of CD133, CD24, and CD44 was analyzed in B16 cell subpopulations with different adhesive capacities by flow cytometry.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Beck B, Blanpain C. Nat Rev Cancer. 2013;13:727–738. - PubMed
    2. Nguyen LV, Vanner R, Dirks P, Eaves CJ. Nat Rev Cancer. 2012;12:133–143. - PubMed
    1. Zhou BB, Zhang H, Damelin M, Geles KG, Grindley JC, Dirks PB. Nat Rev Drug discovery. 2009;8:806–823. - PubMed
    2. McCubrey JA, Steelman LS, Abrams SL, Misaghian N, Chappell WH, Basecke J, Nicoletti F, Libra M, Ligresti G, Stivala F, Maksimovic-Ivanic D, Mijatovic S, Montalto G, Cervello M, Laidler P, Bonati A, Evangelisti C, Cocco L, Martelli AM. Curr Pharm Des. 2012;18:1784–1795. - PubMed
    3. Ishizawa K, Rasheed ZA, Karisch R, Wang Q, Kowalski J, Susky E, Pereira K, Karamboulas C, Moghal N, Rajeshkumar NV, Hidalgo M, Tsao M, Ailles L, Waddell TK, Maitra A, Neel BG, Matsui W. Cell stem cell. 2010;7:279–282. - PMC - PubMed
    1. Tirino V, Desiderio V, Paino F, Papaccio G, De Rosa M. Methods Mol Biol. 2012;879:513–529. - PubMed
    1. Gupta PB, Fillmore CM, Jiang G, Shapira SD, Tao K, Kuperwasser C, Lander ES. Cell. 2011;146:633–644. - PubMed
    1. Farahani E, Patra HK, Jangamreddy JR, Rashedi I, Kawalec M, Rao Pariti RK, Batakis P, Wiechec E. Carcinogenesis. 2014;35:747–759. - PubMed
    2. Zetter BR. Semin Cancer Biol. 1993;4:219–229. - PubMed

Publication types

LinkOut - more resources