Prostaglandin E2 supports growth of chicken embryo intestinal organoids in Matrigel matrix. (May 2012)

• Record Type: Journal Article
• Authors: Pierzchalska, Malgorzata; Grabacka, Maja; Michalik, Marta; Zyla, Krzysztof; Pierzchalski, Piotr
• Is Part Of: Biotechniques. Volume 52:Number 5(2012)
• Page Start: 307
• Page End: 315
• Publication Date: 2012-05
• Subjects: intestinal epithelial cells -- intestinal organoids -- SOX-9 -- 3-D culture -- Matrigel -- chicken embryo; Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/0000113851
• Languages: English
• ISSNs: 0736-6205

Abstract

Investigating intestinal physiology in vitro remains challenging due to the lack of an effective primary enterocyte culture system. Recently developed protocols for growing organoids containing crypts and villus from adult mouse intestinal epithelium in Matrigel present an attractive alternative to the classical techniques. However, these approaches require the use of sophisticated and expensive serum-free medium supplemented with epithelial growth factor (EGF), Wnt agonist (R-spondin 1), and bone morphogenetic protein inhibitor (Noggin) in high concentrations. Here we demonstrate that is possible to use an isolated chicken embryonic intestinal epithelium to create such an organoid culture. Structures formed in Matrigel matrix in the first two days following isolation survive and enlarge during ensuing weeks. They have the appearance of empty spheres and comprise cells expressing cytokeratin (an epithelial cell marker), villin (a marker of enterocytes), and Sox-9 (a transcription factor characteristic of progenitors and stem cells of intestinal crypts). With chicken embryonic tissue as a source of organoids, prostaglandin E2 is as effective as R-spondin 1 and Noggin in promoting sustained growth and survival of epithelial spheroids.