A Functional Assay for Putative Mouse and Human Definitive Endoderm using Chick Whole-Embryo CulturesMartina Johannesson1, Henrik Semb2, Palle Serup2 and Mattias Hansson1*
- *Corresponding Author:
- Mattias Hansson
Hagedorn Research Institute
Niels Steensens Vej 6
DK-2820 Gentofte, Denmark
Tel: +45 4442 9137
E-mail: [email protected]
Received date July 20, 2012; Accepted date September 12, 2012; Published date September 14, 2012
Citation: Johannesson M, Semb H, Serup P, Hansson M (2012) A Functional Assay for Putative Mouse and Human Definitive Endoderm using Chick Whole- Embryo Cultures. J Stem Cell Res Ther 2:128. doi:10.4172/2157-7633.1000128
Copyright: © 2012 Johannesson M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Introduction: Embryonic stem cells (ESCs) represent a prospective cell source for treating degenerative diseases such as diabetes. Several studies have addressed the generation of definitive endoderm (DE) from this cell source by attempting to recapitulate the signaling events occurring during embryogenesis. However, the subsequent differentiation of DE has failed to generate functional insulin-producing beta cells. To assure that we have the correct starting material, we need to fully characterize ESC-derived DE, by assessing whether the cells are functionally equivalent to the in vivo counterpart. Thus, the purpose of this study is to describe a method whereby the in vivo functionality of DE derived from ESCs can be assessed.
Methods: By directed differentiation, putative DE was derived from human and mouse ESCs. This putative DE was subsequently transplanted into the endoderm of chick embryos to determine any occurrence of integration. Putative DE was analyzed by gene and protein expression prior to transplantation and 48 h post transplantation.
Results: Putative DE, derived from mouse and human ESCs, was successfully integrated within the chick endoderm. Endoderm-specific genes were expressed in the putative DE prior to integration and endoderm-specific proteins were assessed 48 h post transplantation.
Conclusions: We describe the detailed methodological procedure for transplanting putative DE derived from ESCs, and the subsequent analysis of the migration and development of the grafted cells. Our result show that putative DE integrates with the chick endoderm and participate in the development of the chicken gut, indicating the generation of functional DE from ESCs. This functional assay can be used to assess the generation of functional DE derived from both human and mouse ESCs and provides a valuable tool for cell characterization. This is an important initial step in the differentiation process towards fully functional beta cells.