• 제목/요약/키워드: Glycerol analysis

검색결과 168건 처리시간 0.026초

영유아용 조제분유의 위치별 지방산 및 Triacylglycerols의 정성 분석 (Determination of Positional Fatty Acid and Triacylglycerol Compositions of Selected Infant Formulas)

  • 손정매;이정희;홍순택;이경수;박혜경;권광일;이기택
    • 한국식품영양과학회지
    • /
    • 제40권9호
    • /
    • pp.1256-1264
    • /
    • 2011
  • 시중에 유통되고 있는 조제분유의 지방산 및 TAG 조성을 알아보기 위하여 국내외 조제분유 총 11 품목을 선정하였고, Folch법을 이용하여 지방을 추출한 다음 총 지방산과 sn-2 위치의 지방산 및 silver ion HPLC를 이용한 단일불포화-포화-단일불포화 glycerol(MSM)의 조성을 분석하였다. 조제분유의 조지방의 함량은 17.21~23.51%이었고, 특히 K5의 조지방 함량이 23.51%로써 분석된 조제분유들 중에서 가장 높게 나왔으나, K1은 17.21%의 함량을 보이면서 가장 낮게 나타났다. 분석된 조제분유들의 주요 포화지방산은 lauric acid(C12:0), myristic acid(C14:0), palmitic acid(C16:0), stearic acid(C18:0)이었다. Lauric acid의 함량은 K5와 K6에서 19.49%와 23.34%로 분석되어 15%보다 높은 함량을 나타냈다. 한편, 분석된 조제분유의 palmitic acid의 함량은 7.59~21.65%의 범위로써 K2, K3, K6, F1에서 각각 14.64%, 8.29%, 13.47% 및 7.59%로써 낮은 함량을 나타냈다. 불포화 지방산들 중에서 단일불포화지방산(C14:1, myristoleic acid; C16:1, palmitoleic acid; C17:1, heptadecenoic acid; C18:1, oleic acid; C18:1t, elaidic acid)의 함량은 20.23~45.07%로 나타났고, 단일불포화지방산들 중에서 대부분을 구성하고 있는 지방산은 oleic acid(C18:1)이었다. 분석된 조제분유에서 oleic acid의 함량은 19.64~44.64%의 범위였고 총 다가불포화지방산은 19.08~29.67%의 범위였다. Omega-6 다가불포화지방산과 omega-3 다가불포화지방산의 주요한 지방산은 필수지방산인 linoleic acid(C18:2)와 linolenic acid(C18:3, n-3)로 분석된 조제분유의 비율은 10:1~13:1로, 조제분유의 지방산 조성에 대한 지표인 linoleic acid/linolenic acid의 비율인 5:1~15:1 범위 내였다. 분석한 조제분유들 중에서 sn-2 위치에서의 lauric acid 함량은 7.70~52.09%로 나타났고, myristic acid의 함량은 3.15~20.89%로써 분석된 조제분유들은 전반적으로 sn-2 위치에서의 lauric acid 함량이 20% 이상으로 높게 나타났다. 특히, 분석된 조제분유들의 sn-2 위치에 존재하는 palmitic acid의 함량은 1.84~38.74%의 범위로써 K8과 F3이 각각 38.55%와 38.90%의 높은 함량을 나타냈다. 따라서 대부분의 국내외 조제분유들을 구성하는 triacylglycerol(TAG)의 sn-2 위치에서의 palmitic acid 함량이 모유와 비교하였을 때 낮은 경향을 보임을 알 수 있었다. 이러한 조제분유보다 더 나아가 HPLC 분석을 통해서 모유의 주요 TAG 형태라고 할 수 있는 단일불포화-포화-단일불포화 glycerol(MSM)의 함량이 외국의 조제분유인 F3이 가장 높게 나타났다. 따라서 조제분유에 이러한 MSM 형태의 지방 함량을 높여야하며, MSM 형태의 모유대체지 연구를 산업화하려는 노력이 필요하다.

인도네시아 열대작물 오일의 Amberlyst-15 촉매 에스테르화 반응 및 바이오디젤 물성 분석 (Esterification of Indonesia Tropical Crop Oil by Amberlyst-15 and Property Analysis of Biodiesel)

  • 이경호;;이준표;이진석;김덕근
    • 한국응용과학기술학회지
    • /
    • 제36권1호
    • /
    • pp.324-332
    • /
    • 2019
  • 한국과 인도네시아를 포함한 대부분의 국가는 온실가스 감축을 위해 바이오디젤 같은 바이오연료 보급에 대한 강력한 정책을 추진하고 있다. 하지만, 바이오디젤 보급 확대를 위해서는 원료 부족 문제를 먼저 해결해야 한다. 본 연구에서는 원료 공급 안정성을 개선하고 바이오디젤 생산 가격을 낮추기 위해 비식용이면서 동시에 단위면적당 생산성이 높은 인도네시아 열대작물(R. Trisperma) 오일의 바이오디젤 생산 가능성을 조사하였다. 수확기간이 다른 두 종류의 오일은 많은 불순물과 높은 유리지방산 함량을 가지고 있어 효율적인 바이오디젤 생산을 위해, 에스테르화 반응과 전이에스테르화 반응을 실시하였다. 오일은 반응을 진행하기 앞서 여과와 수분제거 과정을 통해 반응의 효율을 높이고자 하였다. 에스테르화 반응은 불균질계 산 촉매인 Amberlyst-15를 사용하였으며, 반응 전 오일들의 산가는 각각 41, 17 mg KOH / g이었으나, 에스테르화 반응 후 3.7, 1.8 mg KOH/g으로 약 90% 이상의 전환율을 보이며 유리지방산 함량을 2%이하로 감소시켰다. 이후 전이에스테르화 반응은 KOH를 염기 촉매로 사용하여 바이오디젤 합성 실험을 진행하였다. 생성된 바이오디젤은 약 93%의 FAME 함량을 나타냈으며, 총 글리세롤의 함량은 0.43%으로 제품 규격(FAME 96.5%, 총 글리세롤 0.24%)에는 미달되었다. 이는 지방산 조성 분석 결과 일반적으로 관찰되지 않는 특이 지방산인 ${\alpha}$-Eleostearic acid가 10.7~33.4% 포함되어 나타나는 특성으로 판단되며, 추가 반응 최적화 및 분리정제 연구 진행으로 연료품질 규격 달성이 필요한 것으로 나타났다. 기존에 활용되지 못하던 비식용 원료로부터 바이오디젤 생산 기술을 확보할 경우 바이오디젤 보급 확대를 위한 안정적 원료 공급에 기여할 것으로 판단된다.

韓.洋方 協診治療를 위한 三叉神經痛의 文獻的 硏究 (Study of literature on the Trigeminal Neuralgia for cooperative treatment of Oriental and Occidental medicine)

  • 성병곤;오춘근;임규상
    • 한방안이비인후피부과학회지
    • /
    • 제13권2호
    • /
    • pp.112-139
    • /
    • 2000
  • I examined and referred to literatures of every generations on the nicknames, causes, herb medications and acupucture treatments of ptosis(上胞下垂). And then the results were obtained as follows. We've compared and analyzed Occidental and Oriental medical causes, symptoms and treatments of Primary trigeminal neuralgia and wanted to get better effects by a cooperative analysis. So the examination and analysis of the recent treatment tendency and reference bibliography show the following results. 1. Trigeminal neuralgia is nerve systematic disease appearing in the distribution scope of trigeminal nerve. It's characterized by extreme pain accompanying with a repeated and simultaneous fit from several seconds to 1-2 minutes. 2. Though there are many hypothesis on the trigeminal neuralgia, but now many doctors agree that when trigeminal nerve is under the local out of sheath conditions resulting from receiving a chronic stimulus, and the nucleus of trigeminal nerve fire, owing to decrease of pain control function and abnormal occurrence of action potential, it would be appeared. 3. The Oriental medical name of trigeminal neuralgia is generally Dootong, Doopoong, Myuntong, Pyundootong, Pyundoopoong, and Myuntong is the nearest in Occidental medicine. 4. The Oriental medical cause of trigeminal neuralgia is usually divided into Wekam and Naesang. The first one is caused by Poonghan, Poongyul, Damhwa and wicked energy enter into the body, the mechanical energy is obstructed and can't move any more, so the pain appears by them. The other cause is the hurt by emotion. And it would be loss of the transportain of liver and obstructed, so result into Kanwulhwahwa, Kanpoongnaedong and the pain appears. 5. There are two methods of curing trigeminal neuralgia. As a medication, primary method is prescribing Carbamazepine and the second is using Phenytoin or Baclofen. And as a operation, Drug injection of trigeminal nerve, Amputation of branches of trigeminal nerve, Retrogasserian glycerol rhizotomy, Radiofrequency gangliolysis, Neurovascular decompression can be used. 6. There are several herb medicines for Trigeminal neuralgia. First, Chungung is good for Hwaejeetong, Keopoongjedam, Hwalhyuljeetong. Second, Jeongal, Jiryong, Okong is used for Sikpoonghekyung, Tongkyungjeetong. Third, Baekjee, Sesin, Cheonma, Manhyungja is efficacious in Sinonhepyo. Sanpoongjeetong. Fourth, for falling of liver's Wulhwa, Yongdamcho, Hyungge, Kukwha can be used. And also Saengjihwang, Hwangkm is good for going down the fever of Yangmyungwiyul and finally, Baekkangjam. Moryu can be effective for Jaumjamyang, Haekyungjitong. The other medicines can be used as assistant analgesics, and it also efficacious. 7. Generally the points of pain on the face and the points of Soyangkyung and Yangmyungkyung is used for Acupuntual therapy, because the two meridians passed on the face. Hakwan. Sabaek, Kwanryo, Keoryo, Hyubkeo, Taeyang, Jeechang, Younghyang, Eoyo, Chanjuk. Yangbaek. Sajukkong. Dooyoo, Kwangsangjum, Sengjang, Poongjee is used for taking near point and Joksamlee, Naejung, Habkok is used for taking distant point. 8. Dansam or Danggui injection which have a effect for Hwalhyulhwaeo, Sokyunghwalak and Vit B1, Vit B2, Vit B12, $2\%$ Hydrochloroprocaine, $1\%$ Lidocaine injection to pain point for local analgesics had so good effect. And external application and moxibustion are used for another treatment. 9. It proved that through mouse model, both Herb medication group and Drug medication group are efficacious for trigeminal neuralgia similarly and also the cooperative medication group shows more effective result than the only drug medication group.

  • PDF

A Clinical Analysis of Secondary Surgery in Trigeminal Neuralgia Patients Who Failed Prior Treatment

  • Kang, Il Ho;Park, Bong Jin;Park, Chang Kyu;Malla, Hridayesh Pratap.;Lee, Sung Ho;Rhee, Bong Arm
    • Journal of Korean Neurosurgical Society
    • /
    • 제59권6호
    • /
    • pp.637-642
    • /
    • 2016
  • Objective : Although many treatment modalities have been introduced for trigeminal neuralgia (TN), the long-term clinical results remain unsatisfactory. It has been particularly challenging to determine an appropriate treatment strategy for patients who have responded poorly to initial therapies. We analyzed the surgical outcomes in TN patients who failed prior treatments. Methods : We performed a retrospective analysis of 37 patients with recurrent or persistent TN symptoms who underwent surgery at our hospital between January 2010 and December 2014. Patients with follow-up data of at least one year were included. The prior treatment modalities of the 37 patients included microvascular decompression (MVD), gamma knife radiosurgery (GKRS), and percutaneous procedures such as radiofrequency rhizotomy (RFR), balloon compression, and glycerol rhizotomy (GR). The mean follow-up period was 69.9 months (range : 16-173). The mean interval between the prior treatment and second surgery was 26 months (range : 7-123). We evaluated the surgical outcomes using the Barrow Neurological Institute (BNI) pain intensity scale. Results : Among the 37 recurrent or persistent TN patients, 22 underwent MVD with partial sensory rhizotomy (PSR), 8 received MVD alone, and 7 had PSR alone. Monitoring of the surgical treatment outcomes via the BNI pain intensity scale revealed 8 (21.6%) patients with a score of I, 13 (35.1%) scoring II, 13 (35.1%) scoring III, and 3 (8.2%) scoring IV at the end of the follow-up period. Overall, 91.8% of patients had good surgical outcomes. With regard to postoperative complications, 1 patient had transient cerebrospinal fluid rhinorrhea (2.7%), another had a subdural hematoma (2.7%), and facial sensory changes were noted in 8 (21.1%) patients after surgery. Conclusion : Surgical interventions, such as MVD and PSR, are safe and very effective treatment modalities in TN patients who failed initial or prior treatments. We presume that the combination of MVD with PSR enabled us to obtain good short- and long-term surgical outcomes. Therefore, aggressive surgical treatment should be considered in patients with recurrent TN despite failure of various treatment modalities.

Effect of Sugar Combination in Tris-buffer on the Viability of Post-thaw Spermatozoa in Canine

  • Yu, D.J.;Jeong, S.R.;Oh, I.S.;Bae, I.H.;Cho, S.G.;Kong, I.K.
    • 한국수정란이식학회:학술대회논문집
    • /
    • 한국수정란이식학회 2002년도 국제심포지엄
    • /
    • pp.90-90
    • /
    • 2002
  • The purpose of this study was to investigate the effect of kind and combination of sugars on the viability and acrosome damage of post-thaw spermatozoa in canine. The extender used was Tris-citric acid extender (Tris-buffer) supplemented with 20% Egg-yolk, 8% glycerol, 1% Equex STM paste, and 70 mM sugars such as monosaccharide (fructose and xylose) and disaccharide(trehalose). To evaluate of sugar combination, the sugars supplemented in Tris-buffer were combined such as single (fructose, xylose, trehalose), two combinations (Fruc+Tre, Fruc+xyl, Tre+xyl) and three combinations (Fruc+Tre+Xyl). The concentration of sperm collected were adjusted of 50${\times}$10$\^$6/ per straw for freezing. The frozen spermatozoa were thawed at 37$^{\circ}C$ for 1 min and then analysis for CASA program in Livestock Improvement Main Center, NACF. The motility of post-thaw spermatozoa in Fruc+Tre was higher than those in fructose, trehalose, xylose, Fru+xyl, Tre+xyl and Fru+Tre+Xyl (79% vs. 63, 66, 70, 71, 74 and 75%). The progressive motility after CASA analysis in Fuc+Tre group was also higher than those in fructose, trehalose, xylose, Fru+xyl, Tre+xyl and Fru+Tre+Xyl (67% vs. 53, 57, 60, 61, 62 and 64%). The acrosome damage of post-thaw spermatozoa stained was not significantly different among treatment groups such as fructose, trehalose, xylose, Fru+Tre, Fru+xyl, Tre+xyl and Freu+tre+xyl (17.7, 18.3, 28.0, 17.0, 19.7, 20.0 and 19.0%). The results indicated that the motility and progressive motility of post-thaw spermatozoa in Fru+Tre group was better, and acrosome normality was not different among all groups. The use of Tris-buffer supplemented with Fru+Tre as sugar for frezing of canine spermatosoa could be better and apply to semen banking and artificial insemination.

  • PDF

銀Ion 및 역상(逆相) HPLC를 이용(利用)한 입체이성체적(立體異性體的) 분석(分析)에 의한 은행종실유(銀杏種實油)의 트리글리세리드의 분자종(分子種)에 관한 연구(硏究) (The Stereospecific Analysis of the Triacylglycerols of Ginkgo Nut Oils by High-Performace Liquid Chromatography(HPLC) in the Silver Ion and Reversed Phase Modes)

  • 조용계;우효경;김훈숙
    • 한국응용과학기술학회지
    • /
    • 제14권1호
    • /
    • pp.61-76
    • /
    • 1997
  • Triacylglycerols of the seeds of Ginkgo biloba have been resolved by high-performace liquid chromatography(HPLC} in the silver-ion and reverse-phase modes. The fatty acids were identified by a combination of capillary gas chromatography and gas-chromatography /mass spectrometry as the methyl and /or picolinyl ester. The main components are $C_{18:2{\omega}6}$(39.0mol%), $C_{18:1{\omega}7}$(asclepic acid 21.5mol%), and $C_{18:1{\omega}9}$(oleic acid, 13.8mol%). Considerable amounts of unusual acid such as $C_{20:3{\Delta}^{5,11,14}$ (5.7mol%), $C_{18:2{\Delta}^{5,9}$(2.8mol%), and $C_{18:3}{\Delta}^{5,9,12}$(1.6mol%), were checked. In addition, an anteiso-branched fatty acid, 14-methylhexadecanoic acid, was also present as a minor component(0.9 mol%). The triacylglycerols were separated into 17 fractions by reverse-phase HPLC, and the fractionation was achieved according to the partition numnber(PN) in which a ${\Delta}^5$-non methylene interrupted double bond($^5$-NMDB) showed different behaviour from a methylene interrupted double bond in a molecule with a given cahinlength. Silver-ion HPLC exhibited excellent resolution in which fractions(23 fractions) were resolved on the basis of the number and configuration of double bonds. In this instance, the strength of interaction of a ${\Delta}^5$-NMDB system with silver ions seemed to be weaker than a methylene interrupted double bond system. The principal triacylglycerol species are as follows ; $(C_{18:2{\omega}6)2}/C_{18:1{\omega}7}$, $C_{18:1{\omega}9}/C_{18:1{\omega}7}/C_{18:2{\omega}6}$, $(C_{18:1{\omega}7)2}/C_{18:2{\omega}6}$, $C_{16:1{\omega}7}/C_{18:1{\omega}9}/C_{20:3}{\Delta}^{5,11,14}$, $C_{16:1{\omega}7}/C_{18:1{\omega}7}/C_{20:3}{\Delta}^{5,11,14}$, $C_{18:1{\omega}9}/C_{18:1{\omega}7}/C_{18:2{\omega}6}$, $C_{18:1{\omega}9}/C_{18:2}{\Delta}^{5,5}/C_{20:3}{\Delta}^{5,11,14}$, $(C_{18:1{\omega}7)2}/C_{18:2{\omega}6}$ and $(C_{18:1{\omega}9)2}/C_{18:2{\omega}6}$, while simple triacylglycerols without $C_{18:2{\omega}6})_3$ were not present. Stereospecific analysis showed that fatty acids with ${\Delta}^5$-NMDB system and saturated chains were predominantly located at the site of sn-3 carbon of glycerol backbones. It is evident that there is asymmetry in the distribution of fatty acids in the TG molecules of Ginkgo nut oils.

Construction of High Sensitive Detection System for Endocrine Disruptors with Yeast n-Alkane-assimilating Yarrowia lipolytica

  • Cho, Eun-Min;Lee, Haeng-Seog;Eom, Chi-Yong;Ohta, Akinori
    • Journal of Microbiology and Biotechnology
    • /
    • 제20권11호
    • /
    • pp.1563-1570
    • /
    • 2010
  • To construct a highly sensitive detection system for endocrine disruptors (EDs), we have compared the activity of promoters with the n-alkane-inducible cytochrome P450 gene (ALK1), isocitrate lyase gene (ICL1), ribosomal protein S7 gene (RPS7), and the translation elongation factor-1${\alpha}$ gene (TEF1) for the heterologous gene in Yarrowia lipolytica. The promoters were introduced into the upstream of the lacZ or hERa reporter genes, respectively, and the activity was evaluated by ${\beta}$-galactosidase assay for lacZ and Western blot analysis for hER${\alpha}$. The expression analysis revealed that the ALK1 and ICL1 promoters were induced by n-decane and by EtOH, respectively. The constitutive promoter of RPS7 and TEF1 showed mostly a high level of expression in the presence of glucose and glycerol, respectively. In particular, the TEF1 promoter showed the highest ${\beta}$-galactosidase activity and a significant signal by Western blotting with the anti-estrogen receptor, compared with the other promoters. Moreover, the detection system was constructed with promoters linked to the upstream of the expression vector for the hER${\alpha}$ gene transformed into the Y. lipolytica with a chromosome-integrated lacZ reporter gene under the control of estrogen response elements (EREs). It was indicated that a combination of pTEF1p-hER${\alpha}$ and CXAU1-2XERE was the most effective system for the $E_2$-dependent induction of the ${\beta}$-galactosidase activity. This system showed the highest ${\beta}$-galactosidase activity at $10^{-6}\;M\;E_2$, and the activity could be detected at even the concentration of $10^{-10}\;M\;E_2$. As a result, we have constructed a strongly sensitive detection system with Y. lipolitica to evaluate recognized/suspected ED chemicals, such as natural/synthetic hormones, pesticides, and commercial chemicals. The results demonstrate the utility, sensitivity, and reproducibility of the system for identifying and characterizing environmental estrogens.

A New Device for Intrauterine Artificial Insemination in the Dog

  • Kong, I.K.;Yu, D.J.;Jeong, S.R.;Oh, I.S.;Yang, C.J.;Cho, S.G.;Bae, I.H.;Oh, D.H.;Kim, H.R.;Cho, S.K.;Park, C.S.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제16권2호
    • /
    • pp.180-184
    • /
    • 2003
  • The intrauterine inseminator (IUI) was developed to provide the method of depositing dog semen into the uterine body instead of the vagina. The IUI consists of a vaginal endoscope, a balloon sheath, and injection catheter. When the endoscope is inserted into the vagina and the balloon expanded with air, the cervical os becomes visible so a injection catheter can be inserted through the cervix for deposition of the frozen-thawed semen. The efficacy of the IUI device was compared to intra-vaginal artificial insemination using semen that had been collected and frozen from pooled sperm-rich fraction of ejaculates collected from two Jindo dog donors. Aliquots of semen were extended with a Tris-egg yolk diluent, centrifuged, the seminal plasma removed, the pellet resuspended with the same diluent, and cooled to $5^{\circ}C$ over a 2 h period. A Tris-egg yolk-glycerol extender was added at $5^{\circ}C$; after 1 h, semen was loaded into 0.5 ml straws, and straws were frozen in LN vapor for 5 min, and immersed in LN for storage. The final sperm concentration for freezing was approximately $100{\times}10^{6}cells/ml$. The straws were thawed at $70^{\circ}C$ for precisely 6 sec, 1.5 ml Tris-egg yolk buffer at $38^{\circ}C$ added, and the 2 ml of thawed semen was used for a single insemination using the IUI device. Each bitch was inseminated at optimal insemination point, which was estimated by vaginal epithelial cells staining and progesterone concentration analysis. Use of the IUI device resulted in 21 of 26 females giving birth to 89 pups ($4.2{\pm}1.6$ pups per litter), while intra-vaginal AI resulted in 6 of 15 females whelping a total of 17 pups ($2.8{\pm}1.2$ pups per litter). We believe the IUI device is easier to use than previously described devices used for intrauterine insemination. In our experience the expansion of the balloon has a calming effect on the bitch that aids the inseminator. These results indicate that the IUI device was able to provide high fertility with 50 million frozen sperm per insemination and two inseminations.

Effects of ice-binding protein from Leucosporidium on the cryopreservation of boar sperm

  • Park, Sang Hyoun;Oh, Keon Bong;Ock, Sun-A;Byun, Sung June;Lee, Hwi-Cheul;Kumar, Suresh;Lee, Sung Gu;Woo, Jae-Seok
    • 한국수정란이식학회지
    • /
    • 제33권3호
    • /
    • pp.185-194
    • /
    • 2018
  • The aim of this study was performed to evaluate the effects of ice-binding protein from the arctic yeast Leucosporidium (LeIBP) supplementation on cryopreservation of boar sperm. The collected semen was diluted ($1.5{\times}10^8/ml$) in lactose egg yolk (LEY) and cooled at $5^{\circ}C$ for 3 h. The cooled semen was then diluted ($1{\times}10^8/ml$) in LeIBP containing LEY with 9% glycerol and maintained at $5^{\circ}C$ for 30 min. The semen was divided into six experimental groups (control, 0.001, 0.005, 0.01, 0.05 and 0.1 mg/ml of LeIBP). The straws were kept on above the liquid nitrogen ($LN_2$) vapors for 20 minutes and then plunged into $LN_2$. After thawing, computer-assisted sperm analysis was used for sperm motility and flow cytometry was performed to assess the viability, acrosome integrity (FITC-PSA/PI), ROS (DCF/PI), lipid peroxidation (BODIPY C11/PI) and apoptosis (Annexin V/PI), respectively. No significant responses were observed for sperm motility. However, sperm viability was significantly increased on 0.05 and 0.1 mg/ml of LeIBP groups compared to control (P < 0.05). In addition, acrosome integrity was significantly increases LeIBP groups (P < 0.05) and both ROS and lipid peroxidation level were lower in all LeIBP groups than those of control (P < 0.05). On the other hand, a significant higher apoptosis rate was observed in 0.05 and 0.1 mg/ml of LeIBP groups compared to control (P < 0.05). It can be assumed that a supplementation of LeIBP in boar sperm freezing extender is an effective method to increase the sperm qualities after cryopreservation.

초자화 동결과 1-단계 융해된 체세포 핵이식란의 직접 이식 기술로 제주흑우 복제소 생산 (Production of Cloned Jeju Black Cattle (Korean Cattle) from SCNT Embryo using Vitrification, One-Step Dilution and Direct Transfer Technique)

  • 김은영;박민지;김재연;박효영;노은지;노은형;송동환;오창언;김영훈;문성호;이동선;고문석;류기중;박세필
    • Reproductive and Developmental Biology
    • /
    • 제35권1호
    • /
    • pp.77-83
    • /
    • 2011
  • One-step dilution and direct transfer would be a practical technique for the field application of frozen embryo. This study was to examine whether Jeju Black Cattle (JBC, Korean Cattle) can be successfully cloned from vitrified and one-tep diluted somatic cell nuclear transfer (SCNT) blastocyst after direct transfer. For vitrification, JBC-SCNT blastocysts were serially exposed in glycerol (G) and ethylene glycol (EG) mixtures [10%, (v/v) G for 5 min., 10% G plus 20% EG (v/v) for 5 min., and 25% G plus 25% EG (v/v) for 30 sec.] which is diluted in 10% FBS added D-PBS. And then SCNT blastocysts were loaded in 0.25 ml mini straw, placed in cold nitrogen vapor for 3 min. and then plunged into $LN_2$. One-step dilution in straw was done in $25^{\circ}C$ water for 1 min, by placing vertically in the state of plugged-end up and down for 0.5 min, respectively. When in vitro developmental capacity of vitrified SCNT blastocyst was examined at 48 h after one-step dilution, hatched rate (56.4%) was slightly lower than that of control group (62.5%). In field trial, when the vitrified-thawed SCNT blastocysts were transferred into uterus of synchronized 5 recipients, a cloned female JBC was delivered by natural birth on day 299 and healthy at present. In addition, when the short tandem repeat marker analysis of the cloned JBC was evaluated, microsatellite loci of 11 numbers was perfectly matched genotype with donor cell (BK94-14). This study suggested that our developed vitrification and one-step dilution technique can be applied effectively on field trial for cloned animal production, which is even no longer in existence.