A Simple Culture Method of Chicken Blastodermal Cells for Germline Transmission

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  • Matsubara Yuko
    Transgenic Animal Research Center, National Institute of Agrobiological Sciences, Japan
  • Hirota Azusa
    Kumagaya Livestock Hygiene Service Center, Japan
  • Sobajima Hideo
    Gifu Prefectural Livestock Research Institute, Japan
  • Kagami Hiroshi
    Faculty of Agriculture, Shinshu University, Japan
  • Tagami Takahiro
    Animal Breeding and Reproduction Research Team, National Institute of Livestock and Grassland Science, Japan
  • Yasue Hiroshi
    National Institute of Agrobiological Sciences, Japan

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Abstract

Introduction of genes into blastodermal cells is one of the procedures for chicken genetic modification in combination with construction of germline chimera. Currently, in vitro cultures of chicken blastodermal cells require feeder cells. In the present study, we employed a simple culture method of chicken blastodermal cells for their germline transmission without using feeder cells. The present study was undertaken to determine whether cultured blastodermal cells of Barred Plymouth Rock (i/i) contained periodic acid-schiff (PAS)-positive materials, alkaline phophatase (ALP), SSEA-1, and EMA-1, which are the markers of pluripotent cells including chicken embryonic stem cells (cESCs). The examination revealed that not all but a large portion of cultured cells contained those markers. Then, the cultured cells (i/i) were injected into White Leghorn recipients (I/I) to generate chimeric chickens. The chimeric chickens thus obtained were crossed with Barred Plymouth Rock, and the cross produced chicks with black feathers (i/i). This observation demonstrated that cultured blastodermal cells were transmitted to germ cells in a chimera. In conclusion, the present culture system provided a tool for genetic modification of chicken.

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