• Biotechnology and Bioprocess Engineering:BBE
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• 제5권5호
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• pp.372-378
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• 2000

The cell culture of Angelica gigas Nakai producing decursin derivatives and immunostimulating polysaccharides was preserved in liquid nitrogen after pre-freezing in a deep freezer at -70$^{\circ}C$ for 480 min. The effects of the cryoprotectant and pretreatment before cooling were investigated to obtain the optimal procedure for cyropreservation. When compared to mannitol, sorbitol, or NaCl with a similar osmotic pressure, 0.7 M sucrose was found to be the best osmoticum for the cryopreservation of A. gigas cells. In the pre-culture medium, the cells in the exponential growth phase showed phase showed the best post-freezing survival after cryopreservation. A mixture of sucrose, glycerol, and DMSO was found to be an effective cryoprotectant and a higher concentration of the cryoprotectant provided better cell viability. When compared with the vitrification, the optimum cryopreservation method proposed in this study would seem to be more effective for the long-term storage of suspension cells. The highest relative cell viability established with the procedure was 89%.

• Clinical and Experimental Reproductive Medicine
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• 제28권4호
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• pp.295-300
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• 2001

Objective : To assess the effects of progesterone and acetyl-L-carnitine used after treated with Isolate�� gradient before semen cryopreservation-thawing on sperm parameters and membrane integrity. Material and Methods : From April 2001 to July 2001, ten normal male partner of couples who were visited in vitro fertilization (IVF) clinics. the semens were treated with $Isolate^{(R)}$ gradient before cryopreservation, spermatozoa was incubated with progesterone (1, 5 and $10{\mu}M$), acetyl-L-carnitine (2.5, 5 and $10{\mu}M$), or both (progesterone, $1{\mu}M$; and acetyl-L-carnitine, $5{\mu}M$) for 30 min. Results: There were no differences in sperm parameters and vital stain among isolate only treated group, progesterone (1, 5 and $10{\mu}M$), acetyl-L-carnitine (2.5, 5 and $10{\mu}M$) and both (progesterone, $1{\mu}M$; and acetyl-L-carnitine, $5{\mu}M$). But, in high concentration of acetyl-L-carnitine ($10{\mu}M$) treated group, sperm parameters and vital stain were decreased. The statistical method was used ANOVA (Kruskal-Wallis test) and p value was <0.01. Conclusions : Neither progesterone nor acetyl-L-carnitine show to be protective effect on the cryodamage assessed by sperm parameters and vital stain (eosin-Y stain) in normal sperm. High concentration of acetyl-L-carnitine ($10{\mu}M$), however, was harmful effect on cryoprevention.

• Clinical and Experimental Reproductive Medicine
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• 제24권2호
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• pp.187-192
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• 1997

The survival rate and motility recovered after cryopreservation of testicular spermatozoa in testicular sperm extraction (TESE)-ICSI program is low. The purpose of this study was to assess the availability and efficiency of mouse empty zona pellucida in cryopreserving human TESE spermatozoa. Mouse empty zonae pellucidae were obtained by extraction of cytoplasm with or without cytochalasin B treatment. Motile sperm from proven-fertile donor and two azoospermic patients after TESE were individually inserted into empty zona pellucida and cryopreserved. Two to five days after cyropreservation, the frozen sperm were thawed and the rates of recovery and motility were observed. The ooplasmic extraction rates of control (N=80) and cytochalasin B treated oocytes (N=80) were 94.0% and 96.2%, respectively (p>0.05). The post-thaw recovery rates of spermatozoa and rates of motility recovery of ejaculate (N=70) and testicular (N=70) sperm were 97.1%, 97.1% and 95.7%, 94.3%, respectively (p>0.05). The results of this study showed that the mouse zone pellucida is useful for cryostorage of single testicular spermatozoa.

• 한국산림과학회지
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• 제107권3호
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• pp.258-265
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• 2018

본 연구는 희귀자생식물인 솜양지꽃의 효율적인 초저온 보존 조건을 탐색하고자 행하였다. 종자의 활력은 PVS2와 PVS3 용액 처리구에서는 약 80% 이상으로 대조구보다 훨씬 높은 활력을 보였다. 종자의 활력은 PVS3가 PVS2 처리보다 높게 나타났다. 종자의 활력은 sucrose처리에도 불구하고 대조구보다 낮았다. PVS2의 60분 처리구와 PVS3의 30분 처리구에서 95%의 발아율을 보였으나 그 외 처리구에서는 발아율이 낮았다. PVS2와 PVS3 처리구에서 배양된 유묘의 생장은 PVS2와 PVS3 용액 30분 처리구를 제외하고는 대조구에 비해 생장이 좋지 않았다. 두 가지 초저온동결보존법 간 유묘 생장에서는 encapsulation법과 vitrification법을 비교한 결과 미세한 차이를 보였으나 통계적으로는 유의성이 없었다. 본 연구는 솜양지꽃의 보존에 도움이 될 것으로 판단된다.