Apical polarity in three-dimensional culture systems: where to now? Page: 4 of 10
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the same in all the ten trillion cells of an individual? Primary cells isolated from tissues are an
ideal starting point for 3D cultures since such cells still contain some of the memories of the
organ. Unfortunately, primary cells are in limited supply, especially from humans. In addition,
they are not easy to manipulate genetically, and individual samples will have considerable
heterogeneity that could interfere with experimental reproducibility. Last but not least we still
have not found a way of making all the relevant cells survive in culture.
As an alternative to primary cultures, cell lines have been developed from both normal and tumor
tissues. Where some fundamental comparisons are made and similarities established between the
primary cells and the respective cell line, then the benefits of studying cell lines are many. These
include such features as tractability, reproducibility, availability and homogeneity, although
these traits need to be recalibrated from time to time in relation to the corresponding primary
cells. Cell lines grown in 3D reduce the complexity of the in vivo state and allow us to
manipulate culture conditions and functions. In addition, they could provide keys to the
information encoded in the tissue architecture.
Many of the 3D culture studies of non-malignant human breast epithelial cells have been
performed using two breast epithelial lines, HMT-3522-S1 (S1)  and MCF10A , both of
which were featured in the initial publication that described the 3D culture assays of human
breast . Both cell lines were isolated from fibrocystic breast tissues, and both form relatively
homo geneous organotypic acinar structures resembling in vivo terminal ductal lobular units
when placed in 3D culture in laminin-rich extracellular matrix (lrECM) (Figure 1). We refer to
any extracellular matrix gel rich in laminin-111 including the commercial products MatrigelTM
and Cultrex as lrECM . Since the initial publication, MCF10A and Si cells have only rarely
both been used in the same study, and so it is welcome that Plachot et at  began their study
with a comparison of these lines so often used separately to study mammary gland function and
polarity. They measured the basoapical polarization of acini formed in 3D lrECM by
immunofluorescent visualization of basal integrins and apical tight-junction proteins. Careful
attention was paid to growing both cell lines according to established protocols for each culture
Plachot et al.  find that both cell lines establish basal polarity; however, the acini-like
structures from MCF10A cells continue to grow with increasingly large apical cavities as a
function of time and acini grown from Si cells do not always contain obvious lumina or only
form small ones. Overall, however, S1 cells have a significantly higher frequency of baso-
apically polarized acini compared with MCF10A. Indeed, the MCF10A cells did not appear to
establish any apical polarity at all under the usual 3D culture conditions. This was demonstrated
by multiple markers, including many tight junction proteins. Plachot et at  then went on to
investigate in more detail the conditions in which Si cells could become apically polarized and
investigated the importance of the media and ECM composition in this process.
The importance of the medium
Despite many studies in the 1950s and 60s that showed the importance of medium composition,
very few scientists pay close attention to the composition of the medium in which they grow
their cells. However, the composition of the media is as consequential as the substratum for how
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Inman, J. L. & Bissell, Mina. Apical polarity in three-dimensional culture systems: where to now?, article, January 21, 2010; Berkeley, California. (digital.library.unt.edu/ark:/67531/metadc830118/m1/4/: accessed February 21, 2019), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.