Figure 1.
Co-culture systems are a powerful tool to test candidate drugs and delivery systems.
The cell cultures are able to mimic in vivo tissues without the ethical and cost issues associated with animal experimentation.
Figure 2.
Optical images of MCF-7 and hFIB co-cultures in different ratios during 10 days of culture.
Co-cultures of MCF-7 to hFIB of ratio: 1∶1 (A1-A5), 1∶3 (B1-B5), 1∶5 (C1-C5), and 3∶1 (D1-D5). Monocultures of MCF-7 (E1-E5) and hFIB (F1-F5) were used as controls. Original magnification 100×.
Figure 3.
Optical images of MCF-7 and hFIB co-cultures after 9 and 10 days.
Co-cultures of MCF-7 and hFIB 1∶1 (A, B) and 1∶5 (C, D). Original magnification 100×.
Figure 4.
Chitosan-Arginine-Histidine and nanoparticles physicochemical characterization.
1H NMR spectra of unmodified CH (A) and CH-H-R (B and D). Zeta potential (C) and SEM analysis (D) of CH-H-R/pDNA nanoparticles, respectively.
Figure 5.
Zeta potential of CH-H-R/pDNA nanoparticles at different pH.
Schematic representation of zeta potential decrease as a function of pH. Data is presented as mean ± s.d.
Figure 6.
CLSM co-cultures at 1∶1 ratio for nanoparticles cellular localization analysis.
Confocal microscopy images of MCF-7 breast cancer cells after 4 h of incubation with nanocarriers (A, B), orthogonal sectioning in xy axis (B1, B2), 3D reconstruction of the cell nucleus (C). Colocalization of the Red and Green channels (D). Colocalization of the Red and Blue channels. Red channel – RITC labelled pDNA/CH-H-R; Green channel – Actin-GFP staining of MCF-7 Blue Channel – Hoechst 33342® nuclear staining. Grey channels: colocalization analysis.
Figure 7.
Confocal laser scanning microscopy images of nanoparticle cellular uptake co-cultures at different ratios.
Co-cultures of MCF-7 to hFIB in ratios of: 1∶1 (A–C), 1∶3 (D–F), 1∶5 (G–I) and 3∶7 (J–L) after 4 h of incubation with nanoparticles. Red channel – RITC-labelled pDNA/CH-H-R nanoparticles; Green channel – Actin-GFP staining of MCF-7; Blue Channel – Hoechst 33342® nuclear staining; Grey Channel: DIC; Merged – Superimposition of all channels.
Figure 8.
Flow cytometry analysis of nanoparticles cellular uptake in MCF-7:hFIB co-cultures.
Representative dot plots of nanoparticle uptake in the different co-culture ratios (A–D). The R2 quadrant was used as a gate for histogram analysis. Representative histograms of nanoparticle uptake in MCF-7 cells (E–H) and hFIB (I–L) populations with different co-culture ratios after 4 h of incubation with RITC-labelled pDNA/CH-H-R nanoparticles.