Oocytes.
Embryonic stem cells (ESCs), derivatives of totipotential cells of early mammalian embryos, have proven to be one of the most powerful tools for studying developmental and stem cell biology. When injected into embryos, ESCs can contribute to tissues derived from all three germ layers and to the germ line. Prior studies have successfully shown that ESCs can recapitulate features of embryonic development by spontaneously forming somatic lineages in culture. More recent studies using differentiating monolayer cultures and embryoid bodies have shown that mouse ESCs can also form germ cells that are capable of undergoing meiosis and forming both male and female gametes. This chapter provides detailed instruction on how to differentiate ESCs in monolayer cultures to derive germ cells and oocyte-like structures and presents standard methodologies for detecting expression of key genetic pathways required for primordial germ cell (PGC) development and oogenesis in vivo. While the full potential of these ESC-derived germ cells and oocyte-like structures remains to be demonstrated, this assay provides a new approach to studying reproductive developmental biology in vitro.
Hübner
Karin
K
Max Planck Institute for Molecular Biomedicine, Cell and Developmental Biology, Muenster, Germany.
Kehler
James
J
Schöler
Hans R
HR
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D002469:Cell Separation / Q000379:methods; D003470:Culture Media; D047108:Embryonic Development; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D005786:Gene Expression Regulation; D008322:Mammals; D051379:Mice; D009865:Oocytes / Q000166:cytology* / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D003470:Culture Media