title: Improved method for ex ovo-cultivation of developing chicken embryos for human stem cell xenografts.
abstract: The characterization of human stem cells for the usability in regenerative medicine is particularly based on investigations regarding their differentiation potential in vivo. In this regard, the chicken embryo model represents an ideal model organism. However, the access to the chicken embryo is only achievable by windowing the eggshell resulting in limited visibility and accessibility in subsequent experiments. On the contrary, ex ovo-culture systems avoid such negative side effects. Here, we present an improved ex ovo-cultivation method enabling the embryos to survive 13 days in vitro. Optimized cultivation of chicken embryos resulted in a normal development regarding their size and weight. Our ex ovo-approach closely resembles the development of chicken embryos in ovo, as demonstrated by properly developed nervous system, bones, and cartilage at expected time points. Finally, we investigated the usability of our method for trans-species transplantation of adult stem cells by injecting human neural crest-derived stem cells into late Hamburger and Hamilton stages (HH26-HH28/E5-E6) of ex ovo-incubated embryos. We demonstrated the integration of human cells allowing experimentally easy investigation of the differentiation potential in the proper developmental context. Taken together, this ex ovo-method supports the prolonged cultivation of properly developing chicken embryos enabling integration studies of xenografted mammalian stem cells at late developmental stages.
authors.affiliation: Molecular Neurobiology, University of Bielefeld, Universitätsstraße 25, 33501 Bielefeld, Germany.
authors.forename: Timo
authors.initials: T
authors.lastname: Schomann
authors.forename: Firas
authors.initials: F
authors.lastname: Qunneis
authors.forename: Darius
authors.initials: D
authors.lastname: Widera
authors.forename: Christian
authors.initials: C
authors.lastname: Kaltschmidt
authors.forename: Barbara
authors.initials: B
authors.lastname: Kaltschmidt
references.citation: J Neurosci. 2006 Mar 22;26(12):3256-68
references.pmid: 16554476
references.citation: Methods Mol Biol. 1999;129:257-69
references.pmid: 10494570
references.citation: J Vis Exp. 2007;(8):306
references.pmid: 18989413
references.citation: Rev Diabet Stud. 2005 Winter;2(4):221-7
references.pmid: 17491698
references.citation: J Vis Exp. 2009 Nov 30;(33):
references.pmid: 19949373
references.citation: Dev Biol. 1974 Dec;41(2):391-4
references.pmid: 4452416
references.citation: Dev Dyn. 1992 Dec;195(4):231-72
references.pmid: 1304821
references.citation: Stem Cells Dev. 2012 Mar 20;21(5):742-56
references.pmid: 22128806
references.citation: Methods Mol Biol. 2000;137:37-42
references.pmid: 10948523
references.citation: Toxicol Lett. 2001 Feb 28;119(2):95-102
references.pmid: 11311570
references.citation: Histol Histopathol. 2010 Feb;25(2):177-87
references.pmid: 20017104
references.citation: Anat Rec. 2002 Jun 1;267(2):177-89
references.pmid: 11997887
references.citation: Pharmacol Toxicol. 1989 Feb;64(2):193-5
references.pmid: 2474162
references.citation: Methods Mol Biol. 2012;872:1-14
references.pmid: 22700400
references.citation: Cell Stem Cell. 2011 Sep 2;9(3):205-18
references.pmid: 21852222
references.citation: Development. 2010 Feb;137(4):585-95
references.pmid: 20110324
references.citation: J Vis Exp. 2010 Oct 23;(44):
references.pmid: 21048670