Novel culture systems for ex vivo development

Abstract

The present invention provides methods for the culture of animal pluripotent stem cells and their differentiated progeny cells, tissues, and organs, and nonhuman animal embryos and fetuses.

1 . A method for culturing mammalian pluripotent stem cells, comprising culturing said cells in an unfertilized telolecithal or eutelolecithal egg.  
   
   
       2 . The method of  claim 1 , wherein the mammalian pluripotent stem cells are human.  
   
   
       3 . The method of  claim 1 , wherein the mammalian pluripotent stem cells are canine or feline.  
   
   
       4 . A method for culturing mammalian pluripotent stem cells, comprising culturing said cells in an fertilized telolecithal or eutelolecithal egg external to the developing non-human animal until the time of the involution of the yolk sac.  
   
   
       5 . The method of  claim 1 , wherein the mammalian pluripotent stem cells are human.  
   
   
       6 . The method of  claim 1 , wherein the mammalian pluripotent stem cells are canine or feline.  
   
   
       7 . A method of culturing a nonhuman mammalian embryo or fetus, comprising culturing said nonhuman mammalian embryo or fetus in an unfertilized telolecithal or eutelolecithal egg.  
   
   
       8 . The method of  claim 4 , wherein the mammalian species is porcine.  
   
   
       9 . A method of reconstituting mammalian cells from chromatin, comprising injecting said chromatin into the oocyte of a telolecithal or eutelolecithal egg, activating the egg, and allowing karyokinesis and cytokinesis to occur.  
   
   
       10 . The method of  claim 6 , wherein the mammalian chromatin is human.  
   
   
       11 . The method of  claim 6 , wherein the oocyte is activated by the introduction and removal of a sperm or sperm head.  
   
   
       12 . A method of culturing mammalian pluripotent stem cells comprising culturing said cells in juxtaposition to the vitelline membrane of an embryonated telolecithal or eutelolecithal egg.  
   
   
       13 . A method of affecting the growth and differentiation of human embryo-derived cells comprising the co-culture of said human embryo-derived cells with cells of a specific pathogen-free species.  
   
   
       14 . The method of  claim 13 , wherein the specific pathogen-free species is avian.  
   
   
       15 . The method of  claim 14 , wherein the human embryo-derived cells are maintained as embryonic stem cell lines by co-culture with chick embryonic fibroblasts feeder cells.  
   
   
       16 . The method of  claim 14 , wherein human embryo-derived cells are differentiated under the inductive effect of avian cells.  
   
   
       17 . The method of  claim 16 , wherein the avian inducer cells are specific pathogen-free.  
   
   
       18 . The method of  claim 16 , wherein the avian inducer cells are differentiated cells.  
   
   
       19 . The method of  claim 18 , wherein the avian inducer cells are mesodermal cells of the aorta-gonadal-mesonephros region and are used to induce hematopoiesis.  
   
   
       20 . The method of  claim 16 , wherein said avian inducer cells are differentiated from avian embryonic stem cells.  
   
   
       21 . The method of  claim 20 , wherein said avian inducer cells derived from avian embryonic stem cells are identified using gene trap markers.