Genetic requirements for normal germ cell formation throughout development

    Project: Research project

    Description

    Defects in the development of human germ cells are common, however, relatively little is known about the
    genes required for normal germ cell formation and function in men and women. Previously, we
    demonstrated the ability of human embryonic stem (ES) cells to differentiate into human germ cells in vitro
    and identified germ line specific genes involved in male and female germ cell development. The studies
    outlined in this proposal aim to enhance the formation of human germ cells from NIH-approved human ES
    cell lines by differentiating ES cells in the presence of Bone Morphogenetic Proteins (BMPs) and enriching
    for VASA-postive germ cells via FACS. Further characterization of human germ cell differentiation will be
    performed throughout development by examining the expression of germ cell specific genes at the mRNA
    level by quantitative RT-PCR and at the protein level by Western Blot analysis and immunohistochemistry.
    Functional analysis of germ cells differentiated from human ES cells will be accomplished by evaluating the
    expression of late germ cell markers, sex specific meiotic genes and meiotic spindle proteins, and by zona
    pellucida binding assays. In addition, the functional properties of both male and female human-ES cell
    derived germ cells will be assessed in vivo using a mouse transplantation system. Moreover, we also
    propose to silence the DAZ/DAZL genes, a family of candidate fertility factors, by RNA interference (RNAi)
    in human ES cells and determine the effects on male and female germ cell formation and function in vitro.
    Approximately 10-15% of couples in our population who are of reproductive age are infertile. This work will
    not only contribute to our understanding of the genetic requirements for normal human germ cell
    development, but it may also provide new therapeutic approaches for the diagnosis and treatment of
    infertility in men and women.
    StatusFinished
    Effective start/end date9/30/079/29/10

    Funding

    • National Institutes of Health: $50,054.00
    • National Institutes of Health: $46,826.00
    • National Institutes of Health: $44,846.00

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    Germ Cells
    Genes
    Bone Morphogenetic Proteins
    Spindle Apparatus
    Aptitude
    Human Development
    Embryonic Stem Cells
    RNA Interference
    Protein Binding
    Fertility
    Cell Differentiation
    Transplantation
    Western Blotting
    Immunohistochemistry

    ASJC

    • Medicine(all)