Related Experiment Videos
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
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.
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.
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.