Glycolysis-Optimized Conditions Enhance Maintenance of Regenerative Integrity in Mouse Spermatogonial Stem Cells during Long-Term Culture

• Published in: Stem cell reports Vol. 8; no. 5; pp. 1430 - 1441
• Main Authors: Helsel, Aileen R., Oatley, Melissa J., Oatley, Jon M.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 09.05.2017; Elsevier
• Subjects: Animals; Cells, Cultured; culture; Glycolysis; Male; Mice; mouse; Primary Cell Culture - methods; Regeneration; Resource; Spermatogenesis; Spermatogonia - cytology; Spermatogonia - metabolism; Spermatogonia - physiology; Spermatogonia - transplantation; spermatogonial stem cell; Stem Cell Transplantation - methods; spermatogenesis; mouse; spermatogonial stem cell; regeneration; culture; glycolysis
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2017.03.004

The application of spermatogonial stem cell (SSC) transplantation for regenerating male fertility requires amplification of SSC number in vitro during which the integrity to re-establish spermatogenesis must be preserved. Conventional conditions supporting proliferation of SSCs from mouse pups have been the basis for developing methodology with adult human cells but are unrefined. We found that the integrity to regenerate spermatogenesis after transplantation declines with advancing time in primary cultures of pup SSCs and that the efficacy of deriving cultures from adult SSCs is limited with conventional conditions. To address these deficiencies, we optimized the culture environment to favor glycolysis as the primary bioenergetics process. In these conditions, regenerative integrity of pup and adult SSCs was significantly improved and the efficiency of establishing primary cultures was 100%. Collectively, these findings suggest that SSCs are primed for conditions favoring glycolytic activity, and matching culture environments to their bioenergetics is critical for maintaining functional integrity. •Regenerative integrity of SSCs declines over time in conventional culture•Glycolysis-optimized (GO) culture improves regenerative integrity of SSCs•GO conditions enhance the long-term culture of SSCs from adult mice In this article, Oatley and colleagues explored the impact of culture conditions on regenerative integrity of mouse spermatogonial stem cells (SSCs). They discovered that long-term maintenance in an environment optimized for utilization of glycolysis improves the capacity to regenerate spermatogenesis after transplantation. Thus, matching culture environments to the bioenergetics of SSCs is critical for maintaining functional integrity.