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A simple and convenient method for preparing chimeric animals from embryonic stem (ES) cells.

Pyapalli Usha Rani1, Jaspal Singh Khillan

  • 1The Wistar Institute, 3601 Spruce Street, Philadelphia, PA 19104, USA.

Transgenic Research
|January 10, 2004
PubMed
Summary

Creating chimeric animals for gene function studies is now cost-effective. This method uses aggregated cryopreserved embryos and green fluorescent protein (GFP) selection for efficient chimeric animal production and gene transmission.

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Area of Science:

  • Developmental Biology
  • Genetics
  • Stem Cell Biology

Background:

  • Gene targeting in embryonic stem (ES) cells is crucial for studying gene function during development.
  • Producing chimeric animals is an effective method for these studies.
  • Existing methods can be costly and time-consuming.

Purpose of the Study:

  • To develop a cost-effective and convenient method for producing chimeric animals.
  • To improve the efficiency of creating gene-targeted chimeric animals using ES cells.

Main Methods:

  • Aggregation of cryopreserved 8-16 cell mouse embryos with ES cells in microwell dishes.
  • Aggregation of freshly isolated morulas with green fluorescent protein (GFP)-positive ES cells.
  • Selection of GFP-fluorescent blastocysts for transfer to pseudo-pregnant mothers.

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Main Results:

  • Chimeric animals were successfully produced with a high degree of ES cell contribution.
  • The generated chimeric animals transmitted the targeted gene to their progeny.
  • The method demonstrated high throughput and cost-effectiveness.

Conclusions:

  • Aggregation with cryopreserved embryos and pre-selection using a visual marker (GFP) is an efficient strategy.
  • This approach facilitates the creation of chimeric animals from gene-targeted ES cells.
  • The developed method offers a practical solution for gene function studies in development and differentiation.