Xenotransplantation of Human Stem Cells into the Chicken Embryo

Abstract

The chicken embryo is a classical animal model for studying normal embryonic and fetal development and for xenotransplantation experiments to study the behavior of cells in a standardized in vivo environment. The main advantages of the chicken embryo include low cost, high accessibility, ease of surgical manipulation and lack of a fully developed immune system. Xenotransplantation into chicken embryos can provide valuable information about cell proliferation, differentiation and behavior, the responses of cells to signals in defined embryonic tissue niches, and tumorigenic potential. Transplanting cells into chicken embryos can also be a step towards transplantation experiments in other animal models. Recently the chicken embryo has been used to evaluate the neurogenic potential of human stem and progenitor cells following implantation into neural anlage [1][2][3][4][5][6] . In this video we document the entire procedure for transplanting human stem cells into the developing central nervous system of the chicken embryo. The procedure starts with incubation of fertilized eggs until embryos of the desired age have developed. The eggshell is then opened, and the embryo contrasted by injecting dye between the embryo and the yolk. Small lesions are made in the neural tube using microsurgery, creating a regenerative site for cell deposition that promotes subsequent integration into the host tissue. We demonstrate injections of human stem cells into such lesions made in the part of the neural tube that forms the hindbrain and the spinal cord, and into the lumen of the part of the neural tube that forms the brain. Systemic injections into extraembryonic veins and arteries are also demonstrated as an alternative way to deliver cells to vascularized tissues including the central nervous system. Finally we show how to remove the embryo from the egg after several days of further development and how to dissect the spinal cord free for subsequent physiological, histological or biochemical analyses. Video LinkThe video component of this article can be found at

Robust derivation of epicardium and its differentiated smooth muscle cell progeny from human pluripotent stem cells

“…We hypothesised that injecting HESC-derived epicardial cells into developing embryos at a stage when the epicardium is formed would promote engraftment in the subepicardial space, eventually contributing to the coronary vasculature. Extra-embryonic intravascular delivery has been previously demonstrated as an efficient method to deliver HESCs to vascularised tissues ( Boulland et al, 2010 ). We therefore administered fluorescent (GFP + or mStrawb + ) HESC-derived epicardial cells into the extra-embryonic circulation of chicken embryos at Hamburger and Hamilton stage (HH) 24, a developmental stage when the epicardium is completely formed and epicardial cells are migrating into the myocardium ( Martinsen, 2005 ).…”

Inhibition of tumor formation and redirected differentiation of glioblastoma cells in a xenotypic embryonic environment

“…The embryos were then harvested at various days posttransplantation. As is typical for transplants of dissociated cells into lesions in the spinal neural tube, not all of the transplanted cells ended up within the neural tube (Sigurjonsson et al, ; Boulland et al, ). On average, approximately half of the transplanted cells remained within the lesion shortly after transplantation (estimated by visual assessment) and of these only a fraction had integrated into the neural tube tissue 1 day posttransplantation, when regeneration was complete.…”

Embryonic Chicken Transplantation is a Promising Model for Studying the Invasive Behavior of Melanoma Cells

“… Make a small nick with a needle in the vitelline membrane overlying the most caudal end of the neural tube. Other procedures use similar processes to prepare chick embryos for grafting experiments ( 29 – 31 ). …”