Establishment of mouse intestinal crypt stem cell-containing, self-sustaining organoids.

<p>(<b>A</b>) Isolation of intestinal crypt cells. The intestinal crypt from the jejunum and ileum of young CD1 mice were harvested as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0093608#s2" target="_blank">Methods</a> (<b><i>a</i></b>). Some intact crypt structures were also retained during the isolation process (<b><i>b</i></b>). (<b>B</b>) The isolated crypt cells were seeded in Matrigel and maintained in the organoid culture medium as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0093608#s2" target="_blank">Methods</a>. The proliferation of crypt cells and organoid formation were apparent at day 3 while budding organoids were readily observed at day 6 after seeding. (<b>C</b>) Representative images of various forms of crypt organoids from the passed crypt culture are shown.