Generation and characterization of porcine induced trophoblast

A Thesis Presented to the Graduate School University of Missouri--Columbia In Partial Fulfillment of the Requirements for the Degree Master of Science.[ACCESS RESTRICTED TO THE UNIVERSITY OF MISSOURI AT AUTHOR'S REQUEST.] Trophoblast stem cells are progenitors of all trophoblast lineages that compose fetal placental portion. Trophoblast stem cells provide an unlimited source of immature trophoblasts as they are capable of self-renewal and multipotency. Furthermore, they can differentiate into mature trophoblast under altered cell culture conditions. These properties are considered favorable for elucidating the mechanism of placental development, eventually leading to an improvement in reproductive efficiency, therapeutics for pregnancy diseases, and perhaps even solving larger enigmatic questions, such as how the diverging placental morphlogies evolved in eutherian mammals. Albeit, only mouse trophoblasts have been established to possess self-renewal and multipotency abilities. Nonetheless, countless scientists have attempted to establish the same cellular properties for stem cells derived from other species. Here, I report that porcine trophoblast cells, designated as induced trophoblast stem cells (iTR), were generated by inducing four reprogramming factors (POU5F1, SOX2, KLF4, MYC) into porcine fetal fibroblast (PFF). Characterization of iTR by examination of gene expression, protein synthesis, ultrastructure, and teratoma formation assay revealed that iTR possess trophoblast features and appear to have self-renewal and multipotency abilities. This is the first study to report the generation of trophoblast cells as a by-product of reprogramming and establishment of porcine trophoblast cells possessing "stemness."Includes bibliographical references (pages 68-77)