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Updated: May 3, 2026
Mouse Sperm Cryopreservation and Recovery using the I·Cryo Kit
Published on: December 12, 2011
Effect of sugar molecules on the cryopreservation of mouse spermatogonial stem cells
Yong-An Lee1, Yong-Hee Kim1, Seung-Jung Ha1
1Department of Animal Science and Technology, Chung-Ang University, Ansung, Gyeonggi-Do, South Korea.
Objective:
To study the influence of sugars and establish a serum-free freezing method for the cryopreservation of spermatogonial stem cells (SSCs).
Design:
Animal study.
Setting:
University laboratory.
Animal(S):
C57BL/6-TgEGFP, C57BL/6 mice.
Intervention(S):
Germ cells enriched from testis cells were frozen using standard freezing medium containing sugars, including monosaccharides, disaccharides, and trisaccharides at 50, 100, and 200 mM, respectively. To study the feasibility of establishing a serum-free freezing method, fetal bovine serum was substituted with knockout serum replacement.
Main Outcome Measure(S):
Freeze-thawed germ cells were evaluated for recovery rate, proliferation capacity, and stem cell activity after transplantation to recipient testes.
Result(S):
Supplementation of freezing medium with 200 mM disaccharide is an effective method for cryopreservation of SSCs. Trehalose is the most effective cryoprotectant among all the sugars tested and only lactose was comparable to trehalose. Our proliferation and transplantation data show that serum-free freezing can be achieved in freezing medium supplemented with 200 mM trehalose, 10% knockout serum replacement, and 10% dimethyl sulfoxide (DMSO) for cryopreservation of SSCs.
Conclusion(S):
These findings raise the possibility of effectively banking frozen SSCs from various species, including humans, in a traditional serum-free medium for germ cell research and male infertility treatments.
