E-ISSN 2218-6050 | ISSN 2226-4485
 

Original Article


The effect of cryopreservation media on the quality of β-thalassemia mouse spermatozoa

Kakanang Buranaamnuay.

Abstract
Background: The mouse model of human diseases is commonly used for biomedical study, including β-thalassemia (β-thal), an inherited hemoglobin disorder. Maintaining the mice strain by natural mating systems is costly and seems impractical, especially during the COVID-19 pandemic. Sperm freezing is a cost-effective solution for β-thal mouse colony management.
Aim: The present study was conducted to determine appropriate cryopreservation media for β-thal mouse spermatozoa to establish a β-thal mouse sperm bank.
Methods: The epididymal spermatozoa of C57BL/6 wild-type (WT) and -globin gene knockout thalassemia (BKO) mice were frozen in four freezing media: I) raffinose-skim milk-monothioglycerol (MTG), II) raffinose-skim milk-glutamine, III) raffinose-egg yolk-glycerol, and IV) egg yolk-TES-Tris. The sperm quality was assessed prior to and following freeze-thawing. Results: Compared with WT counterparts, the viable spermatozoa before freezing exhibiting elevated levels of oxidative stress were significantly greater in BKO (p = 0.01). After thawing, the membrane integrity of BKO spermatozoa preserved in I was significantly lower (p = 0.001). The sperm viability and membrane integrity of BKO males were also inferior when media III and IV were used (p = 0.008 to p = 0.027). The amount of oxidative stress in the spermatozoon of BKO mice was significantly greater when preserved in I, III, and IV (p = 0.002 to p = 0.044). Comparing freezing media, the motility and acrosome integrity of WT and BKO spermatozoa preserved in IV were significantly higher than those in other media (p < 0.001 to p = 0.01). Spermatozoa with the highest mitochondrial membrane potential (MMP) were observed in I in both genotypes (p = 0.012 to p > 0.05). The viability, membrane integrity, and oxidative stress of post-thaw BKO spermatozoa did not significantly differ among freezing solutions.
Conclusion: Irrespective of freezing media, spermatozoa of BKO males are rather more sensitive to cryopreservation than those of WT. Raffinose-skim milk-MTG/glutamine, raffinose-egg yolk-glycerol, and egg yolk-TES-Tris can all be used to preserve BKO mouse spermatozoa. However, with slightly better sperm characteristics, egg yolk-TES-Tris may be a diluent of choice for BKO mouse sperm cryopreservation. The addition of a reducing agent to thawing media is also strongly recommended to efficiently prevent oxidative stress and therefore improve frozen-thawed sperm survival.

Key words: Anemia, Freezing extender, Mouse, Sperm characteristics


 
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Pubmed Style

Buranaamnuay K, . The effect of cryopreservation media on the quality of β-thalassemia mouse spermatozoa. Open Vet J. 2022; 12(5): 602-611. doi:10.5455/OVJ.2022.v12.i5.2


Web Style

Buranaamnuay K, . The effect of cryopreservation media on the quality of β-thalassemia mouse spermatozoa. https://www.openveterinaryjournal.com/?mno=20388 [Access: November 28, 2022]. doi:10.5455/OVJ.2022.v12.i5.2


AMA (American Medical Association) Style

Buranaamnuay K, . The effect of cryopreservation media on the quality of β-thalassemia mouse spermatozoa. Open Vet J. 2022; 12(5): 602-611. doi:10.5455/OVJ.2022.v12.i5.2



Vancouver/ICMJE Style

Buranaamnuay K, . The effect of cryopreservation media on the quality of β-thalassemia mouse spermatozoa. Open Vet J. (2022), [cited November 28, 2022]; 12(5): 602-611. doi:10.5455/OVJ.2022.v12.i5.2



Harvard Style

Buranaamnuay, K. & (2022) The effect of cryopreservation media on the quality of β-thalassemia mouse spermatozoa. Open Vet J, 12 (5), 602-611. doi:10.5455/OVJ.2022.v12.i5.2



Turabian Style

Buranaamnuay, Kakanang, and . 2022. The effect of cryopreservation media on the quality of β-thalassemia mouse spermatozoa. Open Veterinary Journal, 12 (5), 602-611. doi:10.5455/OVJ.2022.v12.i5.2



Chicago Style

Buranaamnuay, Kakanang, and . "The effect of cryopreservation media on the quality of β-thalassemia mouse spermatozoa." Open Veterinary Journal 12 (2022), 602-611. doi:10.5455/OVJ.2022.v12.i5.2



MLA (The Modern Language Association) Style

Buranaamnuay, Kakanang, and . "The effect of cryopreservation media on the quality of β-thalassemia mouse spermatozoa." Open Veterinary Journal 12.5 (2022), 602-611. Print. doi:10.5455/OVJ.2022.v12.i5.2



APA (American Psychological Association) Style

Buranaamnuay, K. & (2022) The effect of cryopreservation media on the quality of β-thalassemia mouse spermatozoa. Open Veterinary Journal, 12 (5), 602-611. doi:10.5455/OVJ.2022.v12.i5.2