Shetty, Anala Vinay Kumar2024-02-092024-02-092020-12https://hdl.handle.net/11299/260612University of Minnesota M.S. thesis.December 2020. Major: Stem Cell Biology. Advisor: Walter Low. 1 computer file (PDF); vi, 86 pages.The ultimate goal in the fields of interspecies chimerism and blastocyst complementation is to overcome the shortage of organs available for transplantation by developing transplantable human organs in relevant host animals such as pigs. In spite of the massive progress in these fields, there are still major hurdles, both technological as well as ethical, that need to be tackled. The current study focuses on developing a 3D ex-vivo Matrigel based system for culturing mouse embryos until the implantation stage. This ex-vivo system can be used to study the development of wild-type and chimeric embryos. This study also demonstrates the strength of single-cell RNA sequencing and analysis in optimizing interspecies chimerism studies. Lastly, this study discusses combining in silico approaches with in vivo and ex-vivo approaches to perform precise stage-matching experiments, to closely analyze development of grafted cells in chimeric embryos, and to discover crucial gene networks for interspecies chimerism.enblastocyst complementationex-vivo cultureinterspecies chimerismmouse embryossingle cell RNA sequencingstem cellsThesis or Dissertation