Browsing by Author "Chiriboga Proano, Jose A"

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    Conditionally immortalizing porcine hematopoietic stem cells using a Hox-B8 retrovirus and characterizing the porcine bone marrow niche
    (2022-05-24) Chiriboga Proano, Jose A; Warren, Amy L; Gilch, Sabine; DeVinney, Rebekah; Galezowski, Angelica; Jenne, Craig
    Studies in the field of veterinary immunology have been hampered by the availability of species-specific research tools that yield more reliable and accurate experimental planning and results. Limited resources exist to study and understand immune cells, such as macrophages, in pigs and other domestic species. Porcine immune cells are often derived in vitro and require isolation from the blood or bone marrow of live animals. The goal of this research project was to conditionally immortalized hematopoietic stem cells in pigs that can produce all immune cells by using an estrogen-controlled Hox-B8 lentivirus. In the presence of estrogen and expression of HoxB8, hematopoietic stem cells would remain in a conditionally immortalized progenitor stage. With removal of estrogen and addition of appropriate cytokines, hematopoietic stem cells would differentiate into a spectrum of common porcine immune cells. To be able to conditionally immortalize the hematopoietic stem cells and prime these cells to differentiate into in vitro, we first had to develop mammalian cell lines capable of expressing and secreting porcine recombinant cytokines - the second aim of this thesis. Methods to isolate porcine bone marrow and create media and conditions to culture bone marrow in the lab were also created in this project, which further allowed us to gain insight into hematopoiesis and the niche microenvironment in pigs.