Exogenous donor-derived hepatogenesis in chimeric hosts

This dissertation investigates advanced genetic engineering techniques and the creation of intra- and interspecies chimeras to explore liver development and the potential for generating donor-derived organs in chimeric hosts. In this dissertation, we begin with a comprehensive review of the literature (Chapter I) regarding histology and anatomy of liver, cardinal functions of the liver, followed by an overview of the current knowledge of embryonic liver development with a specific focus on molecular pathways and genes crucial for hepatogenesis. Specific focus was given to gatekeepr genes such as Foxa, Gata, and Hhex. We also highlight the role of Fah in maintaining liver homeostasis. Additionally, a generalized introduction to advanced genome editing techniques was provided. In Chapter II, we outlined optimized strategies for generating Hhex and Fah knockout embryos using CRISPR/Cas9 and base editing techniques. The outlined methods tested the latest genetic engineering techniques and demonstrated the feasibility of generating successful gene knockouts with improved precision and consistency. This establishes a foundation for creating reliable models to study organogenesis and embryonic development in the context of liver formation. In Chapter III, intra- and interspecies chimeras were then generated by using pluripotent stem cells and blastocyst complementation to assess the integration and functionality of donor-derived liver cells in the host embryos. Our findings reveal both the potential and limitations of intra and intererspecies chimerism in overcoming developmental barriers and achieving successful organogenesis. Building on these results, blastocyst complementation experiments were conducted in a large animal biomedical pig model lacking liver development (ahepatic pigs), successfully generating donor-derived liver tissues. Final Chapter was dedicated to summarize our current findings and a discussion on future directions to enhance the efficiency of generating exogenous organs. Taken together, this body of work is expected to advance our current understanding of the field of regenerative medicine by providing insights into the development of functional, donor-derived organs in chimeric hosts.

Name thesis:degree_name

Ph. D.

Level thesis:degree_level

Doctoral

Discipline thesis:degree_discipline

Animal sciences (MU)

Grantor dc:publisher

University of Missouri--Columbia

Year dc:date.issued

2024

Author dc:creator

• Yeddula, Sai Goutham Reddy

Advisor dc:contributor.advisor

• Telugu, Bhanu

Language dc:language.iso

eng, English

DOI

https://doi.org/10.32469/10355/108948

OAI identifier oai:identifier

oai:mospace.umsystem.edu:10355/108948