• 한국양식학회지
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• 제19권1호
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• pp.52-56
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• 2006

This study was conducted to investigate the effects of three different concentrations (6%, 8% and 10% final volume) of dimethyl sulfoxide (DMSO) on cryopreserved sperm of grass carp (Ctenopharyngodon idellus). Grass carp sperm was suspended in Kurokura extender #2 and equilibrated at $4^{\circ}C$ for 10 min. French straws (0.25 ml) of sperm were frozen from $4^{\circ}C\;to\;-4^{\circ}C$ at a rate of $4^{\circ}C\;min^{-1}$ and then ken $-4^{\circ}C\;to\;-80^{\circ}C$ at a rate of $11^{\circ}C\;min^{-1}$. The straws were kept at $-80^{\circ}C$ for 10 min and finally stored in liquid nitrogen $(-196^{\circ}C)$. The cryopreserved sperm was thawed in a water bath at $40^{\circ}C$ for 30 sec and fertilization, hatching rate and larval malformation were compared with fresh sperm (control). The fertilization rate of post-thawed sperm was comparable (from 88.21% to 94.30%) to that of fresh sperm. However, hatching rate of all frozen sperm were significantly lower (P<0.05) than that of control. Additionally, the larval abnormality rate of frozen sperm was significantly higher than that of fresh sperm. The results indicate that DMSO could affect the quality of cryopreserved sperm of grass carp, and a freezing program and a proper extender composition should be further studied.

• Journal of Korean Neurosurgical Society
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• 제30권5호
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• pp.605-610
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• 2001

Objective : The authors investigated the efficacy of the calcium sulfate(OsteoSet$^{(R)}$ pellets) as an autograft extender when used to perform posterolateral lumbar fusions. Patients and Methods : Twenty patients who underwent lumbar posterolateral arthrodesis for various spinal diseases between October 1999 and March 2000 were evaluated. Arthrodesis was performed by transpedicular screw fixation and bone grafting with a mixture of autograft + calcium sulfate in a 1 : 1 ratio. At time intervals of 1, 2, 3, and 6 months, postoperative radiographs were obtained to review the resorption of calcium sulfate and the evidence of fusion. A modified Lenke scale was used to assess the status of the fusion. Results : At 2 months after operation, the average modified Lenke scale score for the OsteoSet$^{(R)}$ pellets group was 3.8. However at 6 months after operation, the average modified Lenke scale score for the OsteoSet$^{(R)}$ pellets group was 1.8. Resorption of calcium sulfate pellets was revealed in all cases at 6 months after surgery. Conclusion : It is presumed that a combination of calcium sulfate and autograft can play a role as an effective autograft extender in the posterolateral spinal fusion.

• Reproductive and Developmental Biology
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• 제29권3호
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• pp.149-154
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• 2005

The purpose of this study was the analysis of sperm ability in Specific Pathogen Free (SPE) miniature pig for production of bio-organ. The collected semen was diluted with extender and stored at $17^{\circ}C$t for up to 7 days. The semen samples were evaluated at 0, 1, 3, 5, and 7 days of storage for analysis of sperm ability. Sperm ability was evaluated by examining viability, progressive motility, sperm abnormality and intensity of the sperm membrane. Also, the semen was processed according to the convenient freezing method, and frozen-thawed sperm was evaluated by examining viability, capacitation and acrosome reaction using chlortetracycline (CTC) staining. Motility of spermatozoa of SPF miniature pig was significantly (P<0.05) lower on 3 days or later compared to the Duroc, Yorkshire and Landrace in domestic boar. The percentage of abnormal spermatozoa of Landrace were significantly (P<0.05) higher than in SPF miniature pig, Duroc and Yorkshire that had a similar percentage on 5 or 7 days of sperm storage. The percentage of spermatozoa with coiled tail decreased during the storage period but there were no significant difference. On the other hand, viability of frozen-thawed spermatozoa had a significantly (P<0.05) lower in SPF miniature pig than in other domestic boars. CTC patterns had no significant difference, but SPF miniature pig had higher percentage of capacitated spermatozoa and lower percentage of acrosome-reacted it than domestic boars. Therefore, this study suggest that it is necessary to develop the suitable extender and freezing methods methods for the high viable rate and fertilizing ability in vitro.

• 한국동물생명공학회지
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• 제34권2호
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• pp.100-105
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• 2019

The purpose of this study was to evaluate the effect of addition of ethylene glycol, glycerol and sucrose to TCG (Tris, Citric Acid, Glucose, Egg Yolk) and DMSO Frozen. The extender containing Egg yolk concentration (10%, 20%) affects viability and acrosome morphology of rabbit sperm. Sperm viability was then assessed for the freezing extenders TCGD (Tris + Citricacid + Glucose + DMSO), TCGED (Tris + Citricacid + Glucose + Egg yolk + DMSO), TCGGD (Tris + Citricacid + Glucose + Glycerol + DMSO) and TCGSD Tris + Citricacid + Glucose + Sucrose + DMSO) during thawing at 38℃. for 20 seconds, respectively. TCG + 10% egg yolk (viability: 77.0 ± 0.8, NAI: 73.3 ± 0.9) was significantly (sperm viability and normal acrosome interaction (NAI)) higher than TCG + 20% egg yolk (70.7 ± 1.1, 70.0 ± 0.9) in the sperm normalcy analysis according to the yolk concentration. TCGGD (53.4 ± 0.1, 62.3 ± 0.4), TCGSD (61.3 ± 0.0, 67.1 ± 0.1) sperm viability and normal acrosome interaction (NAI) in frozen spermatozoa are TCGD (46.4 ± 2.8 and 56.3 ± 1. 4) and TCGED (23.0 ± 1.1 and 54.6 ± 1.4) extenders was thawed at 38℃ for 20 seconds. According to the results from each frozen bulking agent, sperm membrane integrity by hypotonic swelling test (HOST) analysis in TCGGD (59.8 ± 0.7), TCGSD (59.3 ± 0.5) was significantly high compared to other experimental groups (p < 0.05). In conclusion, these results suggested that TCGGD and TCGSD extenders enhance survivability of rabbit sperm after frozen-thawing.

• Reproductive and Developmental Biology
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• 제40권4호
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• pp.33-37
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• 2016

This study was conducted to evaluate effect of ${\alpha}$-linolenic acid (ALA) and bovine serum albumin (BSA) on viability, acrosome reaction and mitochondrial intact in frozen-thawed boar sperm. The boar semen was collected by gloved-hand method and cryopreserved using freezing extender containing 3 ng/mL ALA and/or $20\;{\mu}g/mL$ BSA. Cryo-preserved boar sperms were thawed in $37^{\circ}C$ water-bath for 45 sec to analysis. Viability, acrosome reaction, and mitochondrial intact were analyzed using flow cytometry. In results, viability of frozen-thawed boar sperm was significantly higher in only ALA+BSA supplement group than control group (p<0.05), whereas there was no difference either in ALA or BSA supplement. However, acrosome reacted sperm in both of live and all sperm population were significantly decreased in all treatment groups than control (p<0.05). Interestingly, mitochondrial intact of boar sperm was enhanced in ALA and ALA+BSA groups compared with control (p<0.05). In this study, we showed that supplementation of ALA and BSA in freezing extender enhanced the sperm viability, mitochondrial intact and decrease acrosomal membrane damage. In conclusion, our findings suggest that quality of frozen-thawed sperm in mammalians could improve by using of ALA and BSA.

• 접착 및 계면
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• 제3권2호
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• pp.1-8
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• 2002

물과의 발포반응에 의해 유츌유를 겔화할 수 있는 폴리우레탄 NCO prepolymer를 제조하기 위하여 폴리올(PTMG 및 GP)과 이소시아네이트(TDI)를 사용하여 NCO prepolymer를 합성하였다. 폴리올 각각의 분자량에 따라 합성한 NCO prepolymer를 이용하여 초기 유출유와 에멀젼된 유출유 그리고 유출유의 종류에 따라 유겔화율을 측정하였다. Bunker B 초기 유출유에 대하여 3관능성 폴리오인 GP1000의 경우 440%의 유겔화율은 나타내었으며, 유출유와 해수가 에멀젼(emulsion)된 상태에서는 2배 정도가 증가한 958%의 유겔화율을 나타내었다. 또한 Bunker B에 비해 점도가 높은 Bunker C의 에멀젼된 상태에서는 1098%의 유겔화율을 나타내었다. 사슬연장제가 투입된 2관능성 폴리올인 PTMG1000의 경우에는 에멀젼된 Bunker B에 대하여 910%의 유겔화율을 나타내었으며, 에멀젼된 Bunker C에 대하여 923%의 유겔화율을 나타내었다. 3관능성 폴리올을 사용하여 제조된 NCO prepolymer의 경우, 형성된 폴리우레탄 겔은 부드럽고 강한 특성을 가져 회수에 용이한 상태를 나타내었다.

• Asian-Australasian Journal of Animal Sciences
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• 제17권11호
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• pp.1501-1508
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• 2004

In this study the effect of extenders and temperatures on sperm viability and fertilizing capacity of boar sperm during long-term storage was investigated. Acrosomal integrity, membrane integrity, motility and hypo-osmotic resistance were evaluated by fluorescence and light microscopy. An in vitro fertilization test was performed to assess the fertilizing capacity of stored spermatozoa. The five diluents tested were ranked according to their ability to maintain sperm functional parameters and Zorlesco (ZO) extender with BSA or with PVA instead of BSA produced the best results. Zorlesco extender substituted with PVA (ZO+PVA) was found to maintain motility both at 15 and 20$^{\circ}C$. within 5 days of storage, but the quality of semen stored at 15$^{\circ}C$ decreased thereafter as compared to semen stored at 20$^{\circ}C$ Semen stored at 5$^{\circ}C$ demonstrated rapid loss of motility already within 24 h. Both fertilization and cleavage of semen stored at 20$^{\circ}C$ in ZO substituted with PVA instead of BSA did not change significantly until day 8 of storage. It is therefore concluded that PVA can be used to substitute for BSA and 20$^{\circ}C$ was more suitable than 15$^{\circ}C$ for boar semen storage, and in vitro fertilizing capacity of spermatozoa was maintained for at least 8 days in ZO+PVA at 20$^{\circ}C$.

• Asian-Australasian Journal of Animal Sciences
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• 제33권11호
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• pp.1763-1769
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• 2020

Objective: This study was conducted to investigate the effect of L-carnitine on the pig semen characteristics during storage. Methods: Spermatozoa samples were examined for spermatozoa quality and then randomly divided into 5 groups: 0 (control), 12.5, 25, 50, and 100 mM L-carnitine. Sperm motility, plasma membrane integrity and antioxidant parameters (total reactive oxygen species, total antioxidant capacity, and malondialdehyde) were evaluated after 0, 3, 5, and 10 day cooled-storage at 17℃. Moreover, ATP content, mitochondria activity as well as sperm-binding and in vitro fertilizing ability of preserved boar sperm were also investigated. Results: Supplementation with 50 mM L-carnitine could effectively maintain boar sperm quality parameters such as sperm motility and membrane integrity. Besides, we found that L-carnitine had positive effects on boar sperm quality mainly through improving antioxidant capacities and enhancing ATP content and mitochondria activity. Interestingly, by assessing the effect of L-carnitine on sperm fertility and developmental potential, we discovered that the extender containing L-carnitine could improve sperm quality and increase the number of sperms bounding to zona pellucida, without improving in vitro fertility and development potential. Conclusion: These findings suggested that the proper addition of L-carnitine to the semen extender improved boar sperm quality during liquid storage at 17℃.

• Asian-Australasian Journal of Animal Sciences
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• 제27권6호
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• pp.791-796
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• 2014

Cryopreservation of epididymal sperm is an effective technique to preserve genetic materials of domestic cats and wild felids when they unexpectedly die. However, this technique inevitably causes detrimental changes of cryopreserved-thawed spermatozoa, for example, by physical damage and excessive oxidative stress. L-carnitine is an antioxidant that has been used to improve sperm motility in humans and domestic animals. This study aimed to investigate the effects of L-carnitine on cat epididymal sperm quality following cryopreservation and thawing. After routine castration, cauda epididymides were collected from 60 cat testes. The epididymal spermatozoa from 3 cauda epididymides were pooled as 1 replicate. Spermatozoa samples (16 replicates) were examined for spermatozoa quality and then randomly divided into 4 groups: 0 mM L-carnitine (control), 12.5 mM, 25 mM and 50 mM L-carnitine. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, plasma membrane integrity, DNA integrity and acrosome integrity were evaluated. The 25 mM L-carnitine significantly improved sperm motility compared with a control group (p<0.05), although this was not significantly different among other concentrations. In conclusion, supplementation of 25 mM L-carnitine in freezing extender improves cauda epididymal spermatozoa motility. The effects of L-carnitine on the levels of oxidative stress during freezing and thawing remains to be examined.

• Reproductive and Developmental Biology
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• 제32권1호
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• pp.59-64
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• 2008

본 연구는 정액의 보존 기간 동안 정액의 질적 변화를 알아보고자 시행하였다. 돼지 정액을 Beltsville Thawing Solution (BTS)에 희석한 후 $17^{\circ}C$에서 5일 동안 보존하였다. 보존 기간 동안 정자의 운동성(%)과 linearity는 3일째부터 유의하게 감소하였으나, 다른 운동 역학 변수에서는 유의적 변화를 나타내지 않았다. 또한, 5일 동안 정액을 보존할 경우 첨체의 온전성에도 변화가 없었다. 그러나 제 4일째부터 첨체 변화가 야기된 정자는 유의적으로 증가하였으나, 수정능 획득이 일어난 정자는 유의적으로 감소하였다. 정액의 보존 기간 동안 첨체의 온전성의 유의적 변화가 없었다. 즉, 보존 기간 3일동안 정자의 질적 운동성 및 첨체 온전성에는 유의적인 변화가 없었으므로 상업용 돼지 액상정액은 $17^{\circ}C$에서 적어도 3일간 수정능력을 만족스럽게 유지함을 보여준다.