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Differential cell adhesion of breast cancer stem cells on biomaterial substrate with nanotopographical cues.

Tan KK, Giam CS, Leow MY, Chan CW, Yim EK - J Funct Biomater (2015)

Bottom Line: Topography demonstrated a significant effect on cell adhesion, and the effect was cell type dependent.Flow cytometry analysis showed that 350-nm gratings captured a significantly higher percentage of CD44+CD24- in MCF7.A slightly higher percentage of CD44+CD24-/lowESA+ was captured on the 350-nm gratings, although no significant difference was observed in the CD44+CD24-ESA+ in IDC cells across patterns.

View Article: PubMed Central - PubMed

Affiliation: Mechanobiology Institute, National University of Singapore, T-Lab, #05-01, 5A Engineering Drive 1, Singapore 117411. mbiktkb@nus.edu.sg.

ABSTRACT
Cancer stem cells are speculated to have the capability of self-renewal and re-establishment of tumor heterogeneity, possibly involved in the potential relapse of cancer. CD44+CD24-/lowESA+ cells have been reported to possess tumorigenic properties, and these biomarkers are thought to be highly expressed in breast cancer stem cells. Cell behavior can be influenced by biomolecular and topographical cues in the natural microenvironment. We hypothesized that different cell populations in breast cancer tissue exhibit different adhesion characteristics on substrates with nanotopography. Adhesion characterizations were performed using human mammary epithelial cells (HMEC), breast cancer cell line MCF7 and primary invasive ductal carcinoma (IDC) cells obtained from patients' samples, on micro- and nano-patterned poly-L-lactic acid (PLLA) films. Topography demonstrated a significant effect on cell adhesion, and the effect was cell type dependent. Cells showed elongation morphology on gratings. The CD44+CD24-/lowESA+ subpopulation in MCF7 and IDC cells showed preferential adhesion on 350-nm gratings. Flow cytometry analysis showed that 350-nm gratings captured a significantly higher percentage of CD44+CD24- in MCF7. A slightly higher percentage of CD44+CD24-/lowESA+ was captured on the 350-nm gratings, although no significant difference was observed in the CD44+CD24-ESA+ in IDC cells across patterns. Taken together, the study demonstrated that the cancer stem cell subpopulation could be enriched using different nanopatterns. The enriched population could subsequently aid in the isolation and characterization of cancer stem cells.

No MeSH data available.


Related in: MedlinePlus

(A) Normalized number of IDC cells attached 4 h after seeding on PLLA films (**P < 0.01, ***P < 0.001, mean ± SD, n = 3; except for 1-µm pillars, where n = 1). The IDC cell adhesion behavior normalized to unpatterned control shows preference for nanoscale topography with the 250-nm wells. (B) The normalized number of primary IDC cells cultured after 4 h showed the 250-nm wells as the best topography to capture a significantly higher numbers of CD44+CD24−/lowESA+ cells (***P < 0.001, mean ± SD, n = 3). (C) By immunofluorescence staining and cell counting, the percentage of CD44+CD24−/lowESA+ showed that 350-nm gratings captured a significantly higher percentage of CD44+CD24−/lowESA+ cells (*P < 0.05, mean ± SD, n = 3). (D) Flow cytometry analysis of primary IDC cell culture after 4 h showed that 350-nm gratings captured a higher percentage of CD44+CD24−/low ESA+ cells, but the difference was not significant (mean ± SD, n = 2; except for 1-µm pillars, where n = 1)
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jfb-06-00241-f006: (A) Normalized number of IDC cells attached 4 h after seeding on PLLA films (**P < 0.01, ***P < 0.001, mean ± SD, n = 3; except for 1-µm pillars, where n = 1). The IDC cell adhesion behavior normalized to unpatterned control shows preference for nanoscale topography with the 250-nm wells. (B) The normalized number of primary IDC cells cultured after 4 h showed the 250-nm wells as the best topography to capture a significantly higher numbers of CD44+CD24−/lowESA+ cells (***P < 0.001, mean ± SD, n = 3). (C) By immunofluorescence staining and cell counting, the percentage of CD44+CD24−/lowESA+ showed that 350-nm gratings captured a significantly higher percentage of CD44+CD24−/lowESA+ cells (*P < 0.05, mean ± SD, n = 3). (D) Flow cytometry analysis of primary IDC cell culture after 4 h showed that 350-nm gratings captured a higher percentage of CD44+CD24−/low ESA+ cells, but the difference was not significant (mean ± SD, n = 2; except for 1-µm pillars, where n = 1)

Mentions: Due to the variations among patient samples, primary IDC cell counts varied significantly from one batch to another. The numbers of attached IDC cells on patterned samples were normalized to the number of IDC cells attached on the unpatterned control to facilitate the comparison over three different patient samples. Furthermore, due to the limited cell numbers available from each patient, the cell counting of the total cell number was performed using image analysis of the CellTracker Red- and DAPI-stained cells. From the cell counting, the 250-nm wells showed a significantly higher number of attached primary IDC cells, compared to the 350-nm gratings (P < 0.001) and unpatterned control (Figure 6A, P < 0.01). No significant difference in the normalized cell numbers was observed among the other patterns.


Differential cell adhesion of breast cancer stem cells on biomaterial substrate with nanotopographical cues.

Tan KK, Giam CS, Leow MY, Chan CW, Yim EK - J Funct Biomater (2015)

(A) Normalized number of IDC cells attached 4 h after seeding on PLLA films (**P < 0.01, ***P < 0.001, mean ± SD, n = 3; except for 1-µm pillars, where n = 1). The IDC cell adhesion behavior normalized to unpatterned control shows preference for nanoscale topography with the 250-nm wells. (B) The normalized number of primary IDC cells cultured after 4 h showed the 250-nm wells as the best topography to capture a significantly higher numbers of CD44+CD24−/lowESA+ cells (***P < 0.001, mean ± SD, n = 3). (C) By immunofluorescence staining and cell counting, the percentage of CD44+CD24−/lowESA+ showed that 350-nm gratings captured a significantly higher percentage of CD44+CD24−/lowESA+ cells (*P < 0.05, mean ± SD, n = 3). (D) Flow cytometry analysis of primary IDC cell culture after 4 h showed that 350-nm gratings captured a higher percentage of CD44+CD24−/low ESA+ cells, but the difference was not significant (mean ± SD, n = 2; except for 1-µm pillars, where n = 1)
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jfb-06-00241-f006: (A) Normalized number of IDC cells attached 4 h after seeding on PLLA films (**P < 0.01, ***P < 0.001, mean ± SD, n = 3; except for 1-µm pillars, where n = 1). The IDC cell adhesion behavior normalized to unpatterned control shows preference for nanoscale topography with the 250-nm wells. (B) The normalized number of primary IDC cells cultured after 4 h showed the 250-nm wells as the best topography to capture a significantly higher numbers of CD44+CD24−/lowESA+ cells (***P < 0.001, mean ± SD, n = 3). (C) By immunofluorescence staining and cell counting, the percentage of CD44+CD24−/lowESA+ showed that 350-nm gratings captured a significantly higher percentage of CD44+CD24−/lowESA+ cells (*P < 0.05, mean ± SD, n = 3). (D) Flow cytometry analysis of primary IDC cell culture after 4 h showed that 350-nm gratings captured a higher percentage of CD44+CD24−/low ESA+ cells, but the difference was not significant (mean ± SD, n = 2; except for 1-µm pillars, where n = 1)
Mentions: Due to the variations among patient samples, primary IDC cell counts varied significantly from one batch to another. The numbers of attached IDC cells on patterned samples were normalized to the number of IDC cells attached on the unpatterned control to facilitate the comparison over three different patient samples. Furthermore, due to the limited cell numbers available from each patient, the cell counting of the total cell number was performed using image analysis of the CellTracker Red- and DAPI-stained cells. From the cell counting, the 250-nm wells showed a significantly higher number of attached primary IDC cells, compared to the 350-nm gratings (P < 0.001) and unpatterned control (Figure 6A, P < 0.01). No significant difference in the normalized cell numbers was observed among the other patterns.

Bottom Line: Topography demonstrated a significant effect on cell adhesion, and the effect was cell type dependent.Flow cytometry analysis showed that 350-nm gratings captured a significantly higher percentage of CD44+CD24- in MCF7.A slightly higher percentage of CD44+CD24-/lowESA+ was captured on the 350-nm gratings, although no significant difference was observed in the CD44+CD24-ESA+ in IDC cells across patterns.

View Article: PubMed Central - PubMed

Affiliation: Mechanobiology Institute, National University of Singapore, T-Lab, #05-01, 5A Engineering Drive 1, Singapore 117411. mbiktkb@nus.edu.sg.

ABSTRACT
Cancer stem cells are speculated to have the capability of self-renewal and re-establishment of tumor heterogeneity, possibly involved in the potential relapse of cancer. CD44+CD24-/lowESA+ cells have been reported to possess tumorigenic properties, and these biomarkers are thought to be highly expressed in breast cancer stem cells. Cell behavior can be influenced by biomolecular and topographical cues in the natural microenvironment. We hypothesized that different cell populations in breast cancer tissue exhibit different adhesion characteristics on substrates with nanotopography. Adhesion characterizations were performed using human mammary epithelial cells (HMEC), breast cancer cell line MCF7 and primary invasive ductal carcinoma (IDC) cells obtained from patients' samples, on micro- and nano-patterned poly-L-lactic acid (PLLA) films. Topography demonstrated a significant effect on cell adhesion, and the effect was cell type dependent. Cells showed elongation morphology on gratings. The CD44+CD24-/lowESA+ subpopulation in MCF7 and IDC cells showed preferential adhesion on 350-nm gratings. Flow cytometry analysis showed that 350-nm gratings captured a significantly higher percentage of CD44+CD24- in MCF7. A slightly higher percentage of CD44+CD24-/lowESA+ was captured on the 350-nm gratings, although no significant difference was observed in the CD44+CD24-ESA+ in IDC cells across patterns. Taken together, the study demonstrated that the cancer stem cell subpopulation could be enriched using different nanopatterns. The enriched population could subsequently aid in the isolation and characterization of cancer stem cells.

No MeSH data available.


Related in: MedlinePlus