Generation of BAC Transgenic Epithelial Organoids

作者:Schwank Gerald; Andersson Rolf Amanda; Koo Bon Kyoung; Sasaki Nobuo; Clevers Hans*
来源:PLos One, 2013, 8(10): e76871.
DOI:10.1371/journal.pone.0076871

摘要

Under previously developed culture conditions, mouse and human intestinal epithelia can be cultured and expanded over long periods. These so-called organoids recapitulate the three-dimensional architecture of the gut epithelium, and consist of all major intestinal cell types. One key advantage of these ex vivo cultures is their accessibility to live imaging. So far the establishment of transgenic fluorescent reporter organoids has required the generation of transgenic mice, a laborious and time-consuming process, which cannot be extended to human cultures. Here we present a transfection protocol that enables the generation of recombinant mouse and human reporter organoids using BAC (bacterial artificial chromosome) technology.

  • 出版日期2013-10-18