• 한국가축번식학회지
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• 제15권1호
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• pp.41-47
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• 1991

미성숙 흰쥐에 있어서 과잉배란 유기시 PMSG용량이 체외수정에 의한 수정율과 수정란의 배양에 미치는 영향을 조사하고, 체외수정 및 배양용기로서 plastic mini-straw의 이용효과와 체외수정란의 이식성적을 조사한 바 다음과 같은 결과를 얻었다. 미성숙 흰쥐(체중 65~80g)에게 PMSG를 4, 10, 16 혹은 40IU를 1회 근육주사한 후 72시간에 채란한 난자 중에서 난구세포괴를 가진 정상형태의 난자와 정소상체 미부에서 채취하여 예비배양한 정자로 체외수정시켰다. 체외수정율은 PMSG 용량이 증가될수록 감소하였는데, 즉 4IU에서는 70.8%였으나, 40IU에서는 45.0%로 유의적으로 (P<0.05)저하하였다. 그러나 다정자 수정발생율은 2.3~9.7%로서 PMSG 용량에 따른 유의적인 증가는 없었다. 이 결과는 과량의 PMSG의 투여로 배란된 난자 중에서 일부는 비록 난자가 형태학적으로는 난구세포괴를 가지는 정상적인 난자일지라도 체외수정율의 저하는 정상적인 배란시간보다 조기배란으로 난자의 노화로 인하여 수정에 적합하지 않음을 제시하고 있다. 그리고 plastic mini-straw를 고안하여 straw내에서 체외수정시킨 후 66~72시간까지 배양시험한 결과 2-16와 4-16세포기까지 발달된 배의 비율은 petri dish보다 다소 우수(P<0.05)하였으며, straw용기에서 체외수정된 2세포기의 52개의 배를 7마리의 위임신 흰쥐에게 이식시켰던 바 6개의 배를 이식 받은 한 마리의 수란쥐가 2마리의 새끼를 분만하였다.

• 한국수정란이식학회지
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• 제15권1호
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• pp.95-102
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• 2000

Techniques for manipulation of spermatozoa and oocytes have been widely used for in vitro production(IVP) of Hanwoo. This study was conducted to examine the effects of theophylline and heparin on frozen-thawed Hanwoo sperm for enhancing the efficiency of IVP technique. Oocytes were inseminated with forzen bull semen treated with either theophylline or heparin for examining the effect of each substance on fertilization and subsequent development. More (P<0.05) oocytes formed pronucleus and develop to the morula and blastocyst stages after inseminated with sperm treated with heparin than after inseminated with sperm treated with theophylline. The pregnancy rate after embryo transfer was higher after heparin treatment than after theophylline treatment, but did not differ significantly. There was no significant difference of offspring delivery between two groups. In conculsion, theophylline and heparin can be used for enhancing the efficiency of IVP system for Hanwoo. Considering characteristics of these substance, theophylline may be useful in the artificial insemination system, which requires vigorous sperm motility. While, heparin supporting sperm viability in vitro can be effectively used for improving in vitro-fertilization system.

• 한국가축번식학회지
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• 제21권2호
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• pp.95-102
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• 1997

The studies on the carried out to investigate the effective concentration of cryoprotectant agents and sucrose by one-step straw method of bovine embryos. The follicular oocytes were cultured in TCM-199 medium containing 10 IU/ml PMSG(Sigma, USA), 10 IU/ml hCG(Sigma, USA), 1$\mu\textrm{g}$/ml $\beta$-estradiol(Sigma, USA) and 10% FCS for 24~48 hrs in incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18 hrs with motile capacitated sperm by preincubation of heparin. The bovine embryos following dehydration by cryoprotective agents and various concentrations of sucrose were directly plunged into liquid nitrogen and thawed in 3$0^{\circ}C$ water. Survival and in vitro developmental rate was defined as devellpmental rate on in vitro culture or FDA-test. The results are smmarized as followes : 1. The high in vitro developmental rates of bovine frozen embryos after rapidly thawed in freezing medium was attained 2.0M glycerol, 2.0M DMSO, 1M or 2.0M propanediol. 2. The high in vitro developmental rates of bovine frozen embryos after rapidly thawed in freezing medium was obtained single cryoprotectant(6.7~17.4%) than mixed cryoprotectants(6.7~16.7%). 3. In vitro developmental rate of bovine embryos after rapid frozen-thawing in the freezing medium added 0.25M and 0.50M sucrose were higher cleavage rate than those of sucrose concentration of 0.75M and 1.00M. 4. The freezing methods on in vitro developemental rates of bovine embryos was attained slow freezing method(9.70~15.6%) higher than rapid freezing method(9.4~13.3%).

• 한국자원식물학회지
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• 제19권3호
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• pp.385-391
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• 2006

In vitro-grown axillary buds of Melia aredarach were successfully cryopreserved by vitrification. On the MS medium supplemented with BA 1 mg/L, multiple shoots were developed within $4{\sim}5$ weeks. Plantlets of Melia azedarach were cold-hardened at $10^{\circ}C$ for a 16-hr photo-period for 6 weeks. Excised axillary shoot-tips from hardened plantlets were precultured on a solidified Murashige & Skoog agar medium (MS) supplemented with 0.7 M sucrose for 1 day at $25^{\circ}C$. Axillary shoot-tip meristems wert dehydrated using a highly concentrated vitrification solution (PVS2) for 60 min at $0^{\circ}C$ prior to a direct plunge into liquid nitrogen (LN). The PVS2 vitrification solution consisted of 30% glycerol (w/v), 15% ethylene glycol (w/v), 15% DMSO (w/v) in MS medium containing 0.4M sucrose. After short-term warming in a water bath at $40^{\circ}C$, the meristems were transferred into 2 ml of MS medium containing 1.2M sucrose for 15 min and then planted on solidified MS culture medium. Successfully vitrified and warmed meristems resumed growth within 2 weeks and directly developed shoots without intermediary callus formation. The survival rate of cold-hardened plantlets for 3 and 4 weeks was 90%. We did not find any difference in PCR-band patterns between control and cryopreserved plants. This method appears to be a promising technique for cryopreserving axillary shoot-tips from in vitro-grown plantlets of Medicinal plants.

• Asian-Australasian Journal of Animal Sciences
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• 제25권2호
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• pp.240-247
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• 2012

Dietary supplementation with conventional linted cottonseed hulls (LCSH) is a common practice in livestock production all over the world. However, supplementation with mechanically delinted cottonseed hulls (DCSH) and cottonseed linter residue (CLR) is uncommon. Cottonseed by-products, including LCSH, DCSH and CLR, were assessed by chemical analysis, an in situ nylon bag technique, an in vitro cumulative gas production technique and in vitro enzyme procedure. The crude protein (CP) content of CLR (302 g/kg dry matter (DM)) was approximately 3 times that of LCSH and 5 times that of DCSH. The crude fat content was approximately 3 times higher in CLR (269 g/kg DM) than in LCSH and 4 times higher than in DCSH. Neutral detergent fibre (311 g/kg DM) and acid detergent fibre (243 g/kg DM) contents of CLR were less than half those of DCSH or LCSH. Metabolisable energy, estimated by in vitro gas production and chemical analyses, ranked as follows: CLR (12.69 kJ/kg DM)>LCSH (7.32 kJ/kg DM)>DCSH (5.82 kJ/kg DM). The in situ degradation trial showed that the highest values of effective degradability of DM and CP were obtained for CLR (p<0.05). The in vitro disappearance of ruminal DM ranked as follows: CLR>LCSH>DCSH (p<0.05). The lowest digestibility was observed for DCSH with a two-step in vitro digestion procedure (p<0.05). The potential gas production in the batch cultures did not differ for any of the three cottonseed by-product feeds. The highest concentration of total volatile fatty acids was observed in CLR after a 72 h incubation (p<0.05). The molar portions of methane were similar between all three treatments, with an average gas production of 22% (molar). The CLR contained a higher level of CP than did LCSH and DCSH, and CLR fermentation produced more propionate. The DCSH and LCSH had more NDF and ADF, which fermented into greater amounts of acetate.

• Asian-Australasian Journal of Animal Sciences
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• 제12권7호
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• pp.1054-1062
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• 1999

The effects of dry roasting whole faba beans (WFB) and whole lupin seeds (WLS) at 110, 130 or $150{^{\circ}C}$ for 15, 30 or 45 min on rumen (RDCP%), estimated intestinal (IDCP%) and total tract disappearance of CP (TDCP%) and intestinal availability (IARUCP%) of rumen undegraded CP (RUCP%) were determined. The RDCP values were estimated by in sacco technique by incubating nylon bags for 8, 12 and 24 h in the rumen of dairy cows. The IDCP and IARUCP values were estimated using a sequence of ruminal incubation, in vitro incubation in acid-pepsin for 1 h and then in pancreatin for 24 h of three-step in vitro procedure technique. Dry roasting at 130 and $150^{\circ}C$ decreased RDCP with correspondingly increasing IDCP. The IDCP value generally increased from 12.3(raw) to 8.6, 14.8 and 39.6% (WFB) and from 28.3 (raw) to 33.7, 36.2 and 56.2% (WLS) at 8 h rumen incubation; from 2.9 (raw) to 2.9, 4.6 and 23.3% (WFB) and from 19.6 (raw) to 19.0, 24.0 and 46.6% (WLS) at 12 h rumen incubation; from 1.3 (raw) to 1.9, 1.7 and 11.0% (WFB) and from 4.4 (raw) to 4.2, 10.7 and 36.7% (WLS) at 12 h rumen incubation as the temperatures rose to 110, 130 and $150{^{\circ}C}$ respectively. The TDCP values were always high and increased by time in the rumen, the average values of which were 97.9, 96.6; 99.2, 96.9 and 99.6, 98.7% for WFB and WLS, respectively, at 8, 12 and 24 h rumen incubation. But within the same retention time, TDCP was generally unchanged. The average IARUCP increased from 87.3 (raw) to 87.4, 88.7 and 92.0% (WFB); from 87.6 (raw) to 88.9, 91.5 and 93.0% (WLS) at roasting temperatures of 110, 130 and $150{^{\circ}C}$, respectively. It was concluded that dry roasting can shift the digestion of CP from rumen to the lower gastrointestinal tract without depressing the digestion of RUCP. The best processing condition in this study was dry roasting at $150{^{\circ}C}$ for 45 min in terms of effects on the disappearances and availability of CP. Research data on intestinal availability of individual amino acids need to be further investigated.

• Asian-Australasian Journal of Animal Sciences
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• 제8권5호
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• pp.433-441
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• 1995

A series of experiments was conducted to determine the influence of various pepsin-HCL pretreatment factor, hereby the factors of duration of washing for the retrieved bags, inherent to the mobile nylon bag technique (MNBT), on apparent ileal digestibility of crude protein (AIDCP) and apparent ileal digestibility of dry matter (AIDDM). At last, the AIDCP and apparent ileal digestibility of amino acids (AIDAA) in maize, barley, wheat, rapeseed meal, cottonseed meal and three mixed diets were determined with the MNBT and ileo-rectal anastomis pigs (IRAT). For the MNBT techniques, bag measuring $25{\times}40$ MM and containing 0.75 g feedstuff samples, after pre-digestion in vitro, were introduced into the ileo-rectal anastomis pigs (IRAT) gastrointestinal tract through a duodenal cannula and recovered in the ileal digesta between 6 and 12 h. later. 1. The apparent ileal digestibility of dry matter (AIDDM) and crude protein (AIDCP) of the tested samples, with the exception of fish meal, determined by MNBT were not affected by the different pepsin-HCL pretreatment times in vitro between 2.5 h. and 4 h. 2. There was no significant (p > 0.05) difference of the AIDCP and AIDDM of maize determined by the MNBT among different pepsin concentration (0.03%, 0.07% and 0.1 %) treatment in vitro. 3. The AIDCP determined with the MNBT was affected by the washed and unwashed recovered bags from the ileal digesta. 4. The AIDCP and AID amino acids (AIDAA) of maize, barley, wheat, rapeseed meal, soya-bean meal, cottonseed meal and three mixed diets from the MNBT, with a solution of 0.01N HCL (PH 2) and 0.1% of pepsin concentration, a pepsin-HCL pretreatment time in vitro or 4h. and a washing time of the recovered bag from the ileal digesta compared well with those from the IRAT. The linear regression analysis showed a significant correlation (p < 0.01) of AIDCP and AIDDA between the IRAT and MNBT.

• Corrosion Science and Technology
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• 제15권2호
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• pp.43-53
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• 2016

In this study, electrochemical impedance spectroscopy (EIS) was used to examine the changes in the electrochemical properties of biodegradable pure magnesium implanted into Sprague-Dawley rats for three days. The in vivo test results were compared with those of the in vitro tests carried out in Hank's, dilute saline and simulated body fluid (SBF) solutions. The in vitro corrosion rates were 20~1700 fold higher, as compared to the in vivo corrosion rates. This discrepancy is caused by biomolecule adsorption on the surface, which prevents the transport of water into the magnesium surface on in vivo testing. Among the in vitro experimental conditions, the corrosion rate in SBF solution had the least difference from the in vivo implanted specimen.

• 한국미생물·생명공학회지
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• 제48권4호
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• pp.423-428
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• 2020

In vitro evolution is a powerful technique for the engineering of yeast strains to study cellular mechanisms associated with evolutionary adaptation; strains with desirable traits for industrial processes can also be generated. There are two distinct approaches to generate evolved strains in vitro: the sequential transfer of cells in the stationary phase into fresh medium or the continuous growth of cells in a chemostat bioreactor via the constant supply of fresh medium. In culture, evolutionary forces drive diverse adaptive mechanisms within the cell to overcome environmental or intracellular stressors. Especially, this engineering strategy has expanded to the field of human cell lines; the understanding of such adaptive mechanisms provides promising targets for the treatment of human genetic diseases and cancer. Therefore, this technology has the potential to generate numerous industrial, medical, and academic applications.

• 한국가축번식학회지
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• 제21권2호
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• pp.103-109
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• 1997

The studies on the carried out to investigate to determine the optimum thawing temperature and equilibration time and 1 step straw method of frozen bovine embryos. The follicular oocytes were cultured in TCM-199 medium containing 10 IU/ml PMSG(Sigma, USA), 10 IU/ml hCG(Sigma, USA), 1 $\mu\textrm{g}$/ml $\beta$-estradiol(Sigma, USA) and 10% FCS for 24~48 hrs in incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18 hrs with motile capacitated sperm by preincubation of heparin. The bovine embryos following dehydration by cryoprotective agents and various concentration of sucrose were directly plunged into liquid nitrogen and thawed in 3$0^{\circ}C$ water. Survival and in vitro developmental rate was defined as developmental rate on in vitro culture or FDA-test. The results are summarized as followes : 1. The equilibration time on in vitro developmental rates of bovine embryos was attained after short period of time(2.5~5 min.) in the freezing medium higher than long period of time (10~20 min.). 2. The temperature thawed at 3$0^{\circ}C$ after rapid freezing of bovine embryos resulted in a significantly higher in vitro developmental rate than did at 2$0^{\circ}C$ and 35$^{\circ}C$. 3. The thawing time on in vitro developmental rates of bovine embryos was attained after short period of time(1~5 min.) in the freezing mediuim higher than long period of time(10min.). 4. The in vitro developmental rates of bovine embryos after rapid frozen-thawing by 1 step straw method in the freezing medium added 1.5M, 2.0M glycerol, DMSO, propanediol and 0.25M, 0.50M, 0.75M, 1.00M sucrose were 12.5~19.4%, 10.0~15.6%, 9.1~13.8% and 6.7~12.9%, respectively.