In vitro generation of zebrafish PGC-like cells

Biol Reprod. 2014 Nov;91(5):114. doi: 10.1095/biolreprod.114.121491. Epub 2014 Sep 24.

Abstract

The possibility of generating primordial germ cells (PGCs) in vitro from noncommitted embryonic cells represents an extremely useful tool in current research. Primordial germ cell in vitro differentiation has been successfully reported in mammals. However, contrary to fish, PGC specification in mammals is an inductive mechanism. This study is the first to date to describe a rapid method for PGC in vitro differentiation in teleosts. Primordial germ cell-like cells were characterized by several lines of evidence, including gene expression, cell complexity, size, and image analysis for the quantification of fluorescence under vasa promoter. Moreover, differentiated cells were able to colonize the genital ridge after transplantation. Differentiation treatments increased the number of PGCs in culture, causing differentiation of cells rather than inducing their proliferation. These results open up the possibility of differentiating genetically modified embryonic cells to PGC-like cells to ensure their transmission to the progeny and could be crucial for an in-depth understanding of germline differentiation in teleosts.

Keywords: embryonic cell differentiation; gamete biology; primordial germ cells; zebrafish.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / pharmacology
  • Animals
  • Animals, Genetically Modified
  • Bone Morphogenetic Protein 4 / pharmacology
  • Cell Count
  • Cell Culture Techniques / methods*
  • Cell Differentiation* / genetics
  • Cells, Cultured
  • Embryo, Nonmammalian
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / physiology*
  • Epidermal Growth Factor / pharmacology
  • Female
  • Gene Expression Regulation, Developmental
  • Genes, Developmental
  • Germ Cells / cytology
  • Germ Cells / physiology*
  • Gonadotropins / pharmacology
  • Male
  • Tretinoin / pharmacology
  • Zebrafish Proteins / pharmacology
  • Zebrafish* / embryology

Substances

  • Amino Acids
  • Bone Morphogenetic Protein 4
  • Gonadotropins
  • Zebrafish Proteins
  • bmp4 protein, zebrafish
  • Tretinoin
  • Epidermal Growth Factor