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Self renewal, expansion, and transfection of rat spermatogonial stem cells in culture
Open accessAbstract
The use of a transgenic line of rats that express enhanced GFP (EGFP) exclusively in the germ line has allowed a separation of feeder layers and contaminating testis somatic cells from germ cells and the identification of a set of spermatogonial stem cell marker transcripts. With these molecular markers as a guide, we have now devised culture conditions where rat spermatogonial stem cells renew and proliferate in culture with a doubling time between 3 and 4 days. The marker transcripts increase in relative abundance as a function of time in culture, and the stem cells retain competency to colonize and develop into spermatids after transplantation to the testes of recipient rats. The cells also remain euploid after at least 12 passages. Cell lines could be isolated and cryopreserved and, upon subsequent thawing, continue to self renew. Transfection of the spermatogonial stem cells with a plasmid containing the neomycin phosphotransferase ( neo ) selectable marker resulted in selection of G418-resistant cell lines that effectively colonize recipient testes, suggesting that gene targeting is now feasible in the rat.
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Citations
Cited by
The Germline Stem Cell Niche Unit in Mammalian Testes
Oatley, J. M., & Brinster, R. L. (2012) · Physiological Reviews · doi:10.1152/physrev.00025.2011 · Cited by 513
Progress and prospects in rat genetics: a community view
Aitman, T. J., Critser, J. K., Cuppen, E., Dominiczak, A., Fernandez-Suarez, X. M., Flint, J., Gauguier, D., Geurts, A. M., Gould, M., Harris, P. C., et al. (2008) · Nature Genetics · doi:10.1038/ng.147 · Cited by 291
From engineering to editing the rat genome
Meek, S., Mashimo, T., & Burdon, T. (2017) · Mammalian Genome · doi:10.1007/s00335-017-9705-8 · Cited by 63
2015 Guidelines for establishing genetically modified rat models for cardiovascular research
Flister, M. J., Prokop, J. W., Lazar, J., Shimoyama, M., Dwinell, M., & Geurts, A. (2015) · Journal of Cardiovascular Translational Research · doi:10.1007/s12265-015-9626-4 · Cited by 24
Metadata
- Authors
- Hamra, F. K., Chapman, K. M., Nguyen, D. M., Williams-Stephens, A. A., Hammer, R. E., & Garbers, D. L.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 2005-11-17
- Volume
- 102
- Issue
- 48
- Pages
- 17430-17435
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2155882858
- Cited by
- 210
- References
- 0
- FWCI
- 11.90
- Type
- journal article
- Language
- en
- Open access
- green
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- Hamra, F. K., Chapman, K. M., Nguyen, D. M., Williams-Stephens, A. A., Hammer, R. E., & Garbers, D. L. (2005). Self renewal, expansion, and transfection of rat spermatogonial stem cells in culture. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.0508780102
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