Carmen Crutchfield

For many end-stage diseases, organ transplantation is a final resort when other treatments have been rendered ineffective. Lack of organ availability has forced scientists to investigate novel sources of organs for transplantation in humans. One area of research is interspecies chimerism and blastocyst complementation, which aims to eventually use animals as bio incubators to grow complete human organs within the body of a physiologically similar host species. The objective of this experiment was to evaluate to what extent and to which lineages rat donor cells contributed to in rat-mouse HHEX chimeras. The methods included immunohistochemical (IHC) staining of wild-type mouse embryos for liver-specific markers with anti-HHEX and anti-Albumin antibodies. Performing IHC protocol on wild-type embryos was done to ensure primary antibodies worked as intended before repeating the protocol with rat-mouse HHEX chimera embryos. The objective of the project was not fulfilled as a result of protocol troubleshooting and time limitations. The results of the research performed displayed a lack of liver-localized fluorescence in wild-type embryonic day 12.5 mouse sections stained with anti-HHEX antibody. This result indicates a need to modify and optimize IHC protocol before progression to staining on chimeric embryos. While orthotopic organ transplantations are increasing in demand worldwide, there is a severe shortage of donors. In the future, identifying a molecular mechanism that increases the efficiency of blastocyst complementation would have a positive impact on the field of research which aims to eventually develop functional human organs in host animals such as pigs.