Media 7: Monitoring airway mucus flow and ciliary activity with optical coherence tomography

Originally published in Biomedical Optics Express on 01 September 2012 (boe-3-9-1978

'Insecta exotica'

Originally published in Biomedical Optics Express on 01 September 2012 (boe-3-9-1978

Media 2: Monitoring airway mucus flow and ciliary activity with optical coherence tomography

Originally published in Biomedical Optics Express on 01 September 2012 (boe-3-9-1978

Media 4: Monitoring airway mucus flow and ciliary activity with optical coherence tomography

Originally published in Biomedical Optics Express on 01 September 2012 (boe-3-9-1978

Media 6: Monitoring airway mucus flow and ciliary activity with optical coherence tomography

Originally published in Biomedical Optics Express on 01 September 2012 (boe-3-9-1978

Media 1: Monitoring airway mucus flow and ciliary activity with optical coherence tomography

Originally published in Biomedical Optics Express on 01 September 2012 (boe-3-9-1978

Longitudinal Study of Mammary Epithelial and Fibroblast Co-Cultures Using Optical Coherence Tomography Reveals Morphological Hallmarks of Pre-Malignancy

<div><p>The human mammary gland is a complex and heterogeneous organ, where the interactions between mammary epithelial cells (MEC) and stromal fibroblasts are known to regulate normal biology and tumorigenesis. We aimed to longitudinally evaluate morphology and size of organoids in 3D co-cultures of normal (MCF10A) or pre-malignant (MCF10DCIS.com) MEC and hTERT-immortalized fibroblasts from reduction mammoplasty (RMF). This co-culture model, based on an isogenic panel of cell lines, can yield insights to understand breast cancer progression. However, 3D cultures pose challenges for quantitative assessment and imaging, especially when the goal is to measure the same organoid structures over time. Using optical coherence tomography (OCT) as a non-invasive method to longitudinally quantify morphological changes, we found that OCT provides excellent visualization of MEC-fibroblast co-cultures as they form ductal acini and remodel over time. Different concentrations of fibroblasts and MEC reflecting reported physiological ratios <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0049148#pone.0049148-Sadlonova1">[1]</a> were evaluated, and we found that larger, hollower, and more aspherical acini were formed only by pre-malignant MEC (MCF10DCIS.com) in the presence of fibroblasts, whereas in comparable conditions, normal MEC (MCF10A) acini remained smaller and less aspherical. The ratio of fibroblast to MEC was also influential in determining organoid phenotypes, with higher concentrations of fibroblasts producing more aspherical structures in MCF10DCIS.com. These findings suggest that stromal-epithelial interactions between fibroblasts and MEC can be modeled <em>in vitro</em>, with OCT imaging as a convenient means of assaying time dependent changes, with the potential for yielding important biological insights about the differences between benign and pre-malignant cells.</p> </div

As phericity of acini.

<p>The minimum asphericity value of 1 indicates a perfect sphere, while less spherical acini have higher asphericity values. Acini comprised of MCF10DCIS.com cells are seen to become increasingly aspherical in the presence of fibroblasts.</p

Acini and lumen size.

<p>Comparison of MCF10A:RMF co-cultures with MCF10DCIS.com:RMF co-cultures shows significantly larger acini and lumen sizes (Student’s t-test, p-value <0.005) at week 2. In MCF10DCIS.com:RMF co-cultures, acini and lumen size are also observed to be highly modulated by the ratio of fibroblasts.</p

3D-OCT image acquisition of the co-cultures, and analysis of the shape and size of acini.

<p>A. 3D-OCT image acquistion: the surface of the gel is aligned near the top of each image, and the depth-resolved light scattering from cells beneath the gel surface is apparent at depths up to ∼1 mm; segmentation of acini to characterize the overall size and the lumen is also shown. B. Temporal changes in acini and lumen sizes analyzed from 3D-OCT images of the co-cultures. C. An example isosurface rendering of an acinus from a 3D-OCT image-stack; slicing of the rendered volume clearly shows the lumen. D. An example 3D rendering of an aspherical acinus.</p