Feeder-free method for culture of bovine and porcine spermatogonial stem cells
AssigneeWashington State University
InventorsJon Michael Oatley, Melissa Joan Oatley
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Abstract
The present invention relates to the production and culture of undifferentiated spermatogonial stem cells that can be maintained long term and are feeder free. The resultant feeder-free populations can be used in any of a number of protocols including the generation of progeny bulls. The present invention includes novel methods required for the successful enrichment of bovine spermatogonial stem cells, novel cell lines and other components used for the same, as well as the resultant stem cell compositions.
Claims19
A method of enriching spermatogonial stem cells (SSCs) from a population of testis-derived cells containing at least one SSC, said method comprising:
- a) providing a culture media that has been contacted with embryonic fibroblast cells and/or somatic cells isolated from the testes;
- or parts thereof, to create a preconditioned media;
- b) introducing to said preconditioned media one or more SSC under conditions suitable for SSC cell maintenance and enrichment.
A method of preconditioning media for enriching spermatogonial stem cells (SSCs) from a population of testis-derived cells containing at least one SSC, said method comprising:
- a) obtaining a serum free media suitable for SSC cell maintenance and enrichment;
- b) obtaining immortalized embryonic fibroblast cells and/or immortalized somatic cells isolated from the testes;
- c) contacting said serum free media with said immortalized cells;
- or parts thereof, to create a preconditioned media;
- d) allowing said cells to reproduce on said media;
- and thereafter
- e) removing said immortalized cells.
Dependencies are inferred from the claim text.