Ovarian follicle cells and constructs for fertility treatment and hormone replacement therapy

An in vitro method for providing a three-layered bioengineered ovarian follicle, comprising:

• (a) providing a double positive population of cells, wherein said double positive population of cells comprises oogonia stem cells and said double positive population of cells is positive for:
• i) DEAD box helicase peptide 4 (DDX4), and
• ii) interferon-induced transmembrane protein 3 (IFITM3);
• (b) inducing meiosis in the double positive population of cells by exposing them to retinoic acid;
• then
• (c) adding granulosa cells to the cells and culturing the cells with follicle stimulating hormone (FSH);
• and then
• (d) adding theca cells and culturing the cells, to thereby provide the three-layered bioengineered ovarian follicle.