The development of immortalized human gonadotrope cells lines that express and secrete follicle-stimulating (FSH) and leutinizing hormones (LH) is proposed. There are currently no immortalized human gonadotropes available for research. Additionally, immortalized gonadotropes would be valuable for the in vitro production of FSH and LH which, when purified, could be used both in the diagnosis and therapeutic treatment of infertility. FSH and LH derived from such cells would have several advantages including more natural glycosylation as compared to their urinary-derived and recombinant counterparts. rFSH has been shown to cause hyperstimulation of the ovaries leading to multiple births and higher risk during pregnancy. The following strategy will be employed to immortalize gonadotropes: Primary cultures of human fetal pituitary cells will be analyzed for hormone secretion and propagated using various growth factors. Proliferating cultures will be retrovirally transfected with one of several v-myc expression vectors. Cells that maintain their differentiated phenotype, secrete hormones, survive past normal senescence, and that show no growth in soft agar will be considered immortalized. Transformed lines may also result from this work, and could serve as a research tools, reservoirs for production of recombinant proteins, or for the generation of native, endogenous FSH and LH. PROPOSED COMMERCIAL APPLICATIONS: There is a need for a safe, alternative source of pituitary FSH and LH for use in the treatment of infertility. Presently, recombinant and urinary- derived FSH are used for infertility treatment. Due to differences in glycosylation compared with the natural isoforms, these hormones may not be the safest or most effective. Immortalized gonadotropes expressing FSH and LH would provide a needed tool for the study of human pituitary biology and a new source for the production of these hormones for use in diagnostics and therapeutics for infertility.
