• 한국가축번식학회지
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• 제9권2호
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• pp.153-157
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• 1985

Ejaculated and epidiymal goat spermatozoa were preserved for 0, 6, 12 adn 18 h, and 0 and 18 h in a semi-aerobic condition at 20-$25^{\circ}C$, and preincubated for 5-6 h in a CO2 incubator in m-KRB solution. Then they were preincubated at different concentrations (3-5, 25-48 and 105-190$\times$107/ml), and ability of penetration into zona-free hamster eggs in vitro was examined. When ejaculated spermatozoa were preincubated in m-KRB solution after presservation for 12 and 18 h, 12 and 29% of zona-free eggs were penetrated, and only 4% of eggs were penetrated by epididymal spermatozoa which were preincubated after preservation for 18 h. When spermatozoa were preincubated at a low concentration, the penetration rates were very low. But when the sperm concentration during preincubation was 25-48 and 105-190$\times$107/ml, the penetration rates increased to about 30%.

• Journal of Microbiology and Biotechnology
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• 제2권2호
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• pp.129-134
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• 1992

Segregational instability of a recombinant plasmid, pDML6, encoding extracellular $\beta$-lactamase in Streptomyces lividans PD6 was characterized by growth kinetic analysis. The quantitative determination of the plasmid harbored in the mycelia was evaluated with mycelia fragmented mechanically, and also with colonies regenerated from protoplasts. Conditions for the formation of protoplasts and regeneration of protoplasts were established. The maximal specific growth rates of the host strain and the plasmid-harboring strain in a chemically defined medium without selection pressure were the same. The probability of plasmid loss from the harbouring cells was higher at higher growth rates. Mathematical models for the prediction of cell growth, substrate uptake, and accumulation of the cloned gene product were developed.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2006년도 The 6th Intermational Symposium on Developmental Biotechnology
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• pp.116-117
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• 2006

• 한국작물학회지
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• 제22권1호
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• pp.41-44
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• 1977

담배에서 약배양에 의한 반수체육종을 실용화하고 저 중촌등의 'D'배지를 이용하여 실험을 하였든바 그 결과를 요약하면 다음과 같다. 1. 배양약중 평균 30% 이상이 반수농식물을 유기하였으며 약당 유기된 식물개체수는 평균 6주이상이었다. 2. F$_1$으로부터 유기된 반수체중 T.M.V 저항성은 이론치와 일치하는 1 : 1의 비를 나타내었다. 3. 반수체식물의 생육조사에서는 예기한바와 같이 모든 측정형질에 대하여 F$_1$의 반수체가 모본의 반수체보다 더큰 변이를 보였다.큰 변이를 보였다.

• Asian-Australasian Journal of Animal Sciences
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• 제11권6호
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• pp.655-660
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• 1998

To produce blastocysts more efficiently, it is required to identity accurately the factors involving embryonic cleavage in the chemically defined medium. Effects of pyruvate, lactate, calcium and protein sources on early cleavage of bovine follicular oocytes were investigated. The percentage of IVF embryos cleaved to ${\geq}$ 2-cell or ${\geq}$ 8-cell was higher in pyruvate (+) and lactate (+) (48 or 14%) than in pyruvate (-) and lactate (-) (22% or 4%), than in pyruvate (+) and lactate (-) (28% or 5%) and than in pyruvate (-) and lactate (+) (40% or 10%). Lactate was more effective than pyruvate during early cleavage of bovine embryos in the chemically defined medium. The percentage of IVF embryo cleaved to ${\geq}$ 2-cell and ${\geq}$ 8-cell in calcium (-) (19 and 6%) was significantly (p < 0.05) lower than in calcium (+) (78 and 45%). The percentage of embryos developed to ${\geq}$ 2-cell showed no significant (p < 0.05) difference among BSA, 1 and 20% FBS (57, 57 and 57%). Also the percentage of A grade embryos developed to ${\geq}$ 2-cell showed no significant (p < 0.05) difference among BSA, 1 and 20% FBS (40, 35 and 28%). The percentage of embryos developed to ${\geq}$ 8-cell showed no significant (p < 0.05) difference among BSA, 1 and 20% FBS (33, 23, and 22%). However, the percentage of A grade embryos developed to ${\geq}$ 8-cell in BSA (24%) was significantly (p < 0.05) higher than in 1 and 20% FBS (13 and 8%). The percentage of embryos developed to ${\geq}$ morula showed no significant (p < 0.05) difference among BSA, 1, 10 and 20% FBS (76, 76, 80 and 68%). The percentage of A grade embryos developed to ${\geq}$ morula in 10% FBS (59%) was significantly (p < 0.05) higher than 20% FBS (43%). The percentage of embryos developed to blastocyst showed no significant (p < 0.05) difference among BSA, 1, 10 and 20% FBS (34, 41, 43 and 32%). However, the percentage of A grade embryos developed to ${\geq}$ blastocysts in 10% FBS (25%) was significantly (p < 0.05) higher than in 20% FBS (8%).

• 한국수정란이식학회지
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• 제25권4호
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• pp.259-262
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• 2010

Optimization of the preimplantation mammalian embryo culture condition was widely focused on refining medium composition under the name of chemically defined media. However, recent research revealed that the alteration of physical environment can be a crucial factor to a successful embryo development. In this study, under the same embryo density, a novel culture device named oil-free micro tube culture (MTC) system was evaluated using porcine parthenogenetic embryos. The activated oocytes were placed into the 0.2 ml thin-wall flat cap PCR tube and cultured to the blastocyst stage. As a preliminary step, embryo density and culture medium volume were optimized under a standard drop culture system. The optimal embryo density range for in vitro culture was 0.5 embryos per ${\mu}l$ in $20\;{\mu}l$ drop (20.5%) and 1.0 embryos per ${\mu}l$ in $10\;{\mu}l$ drop (20.6%). Based on these results, we compared drop culture system and 'MTC' system in terms of the developmental rate to the blastocyst stage. In $20\;{\mu}l$ medium volume, the 'MTC' system showed similar blastocyst formation rate when compared with drop culture system (20.2% versus 20.5%, respectively) while the 'MTC' system showed lower blastocyst formation rate than drop culture system in $10\;{\mu}l$ one (12.7% versus 20.0%, respectively). Therefore the $20\;{\mu}l$ MTC system may be an alternative incubation system for short-distance embryo transport without carrying the $CO_2$ incubator and this provides novel embryo culture device to clinical veterinary embryologists.

• Applied Biological Chemistry
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• 제31권4호
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• pp.382-386
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• 1988

Chemically defined medium을 사용하여 Bacillus sp. $T_2-3$으로 균체외단백질을 생산한 후 이들의 각종 물리화학적 성질을 조사 하였다. 균체외단백질은 전기영동과 gel 여과 결과로 분자량이 약 49,000이 되는 비슷한 2종의 단백질로 구성되어 있었다. 최대흡수파장은 230nm 부근이었으며 aspartic acid를 가장 많이 함유하고 있었다. 또한 균체외단백질의 물에대한 용해도는 55.8%, 0.4% NaOH에 대한 용해도는 28.4%로 albumin과 glutelin계통의 단백질로 생각되었다.

• Clinical and Experimental Reproductive Medicine
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• 제21권1호
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• pp.99-110
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• 1994

One consequence of fertilization in mammals is an increased resistance of the zona pellucida (ZP) to proteases and various chemical reagents. This phenomenon has been called 'zona pellucida hardening' (ZPH), and it is generally accepted that it is caused by the secretory products of cortical granules released by the egg at fertilization. ZP of mouse oocytes maturing in vitro in a chemically defined medium becomes progressively more resistant to solubilization by chymotrypsin ("Spontaneous" ZP hardening). In the present study, it was aimed to find the specificity of spontaneous ZPH in relation to its possible relevance to the cortical reaction and the physiological block to polyspermy. When a maturation inhibitors, cAMP analog(dbcAMP) and phosphodiesterase inhibitor (IBMX) was added to culture medium, it prevent spontaneous ZPH of mouse oocyte during in vitro culture. Thus spontaneous ZPH requires GVBD, since it is prevented by those agents, which inhibit GVBD in vitro. However, culture for 3 hours in the presence of PMA(lOng/ml), a protein kinase C activator, resulted in ZPH without GVBD, thus suggesting that ZPH may be regulated independently apart from the event of GVBD. Pretreatment of mouse oocyte with FBS result in partially inhibitory effect on subsequent spontaneous ZPH. Induction of GVBD in vivo had a inhibitory effect on the spontaneous ZPH, but subsequent spontaneous ZPH. Induction of GVBD in vivo had a inhinbitory effect on the spontaneous ZPII, but had no inhibitory effect on PMA-induced ZPH. Treatment with a microfilament formation blocker(cytochalasin-B) at 1${\mu}g$/ml concentration, resulted in the excellent inhibitory effect on spontaneous ZPH. However cytochalasin-B did not inhibit PMA-induced ZPH. Thus this suggesting that spontaneuse ZPH had a different mechanism from PMA-induced ZPH.

• 한국가축번식학회지
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• 제10권1호
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• pp.109-120
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• 1986

This study was conducted to define the effect of addition of lysolecithin (LC) and 20% v/v rabbit serum to sperm preincubation medium on the induction of acrosome reaction (AR) an fertilizing ability in vitro of LG-added sperm. Ejaculated rabbit sperm from New Zealand White buck was washed once by centrifugation, then preincubated for 2 or 4 hrs in a chemically defined medium (DM), DM plus 20% rabbit serum or BSA-free DM plus 20% rabbit serum at 37$^{\circ}C$ water bath or CO2 incubator. At the end of preincubation LC was added to the preincubated sperm, which was stained at 0.5 to 4 hr later and examined for AR and sperm motility. For in vitro fertilization, gametes were coincubated in DM up to 24 hrs and thereafter fertilized embryos were incubated in BSM -II up to 48 hrs. Addition of LC to 4-hr preincubated sperm was more effective for the AR and sperm motility than that to 2-hr preincubated sperm and optimal concentration of LC for AR was about 80${\mu}$g/ml. A significant increase in AR occured from 20 to 30 min. after addition of 80 to 100${\mu}$g/ml in 4-hr preincubated sperm. BSA-free DM plus 20% rabbit serum showed a higher AR and sperm motility than those of DM plus 20% rabbit serum in LC-added sperm after 4-hr preincubation. The incidence of AR after 4-hr preincubation and at 30 min after 60${\mu}$g/ml LC addition varied greatly among individual bucks. Sixty ${\mu}$g/ml LC-added sperm showed a slight high cleavage rate over control levels, but 100${\mu}$g/ml LC-added sperm showed lower cleavage rate rather than 60${\mu}$g/ml LC. It is concluded that optimal concentration of LC for high AR induction and sperm motility in 4-hr preincubated sperm was about 80${\mu}$g/ml, but 60${\mu}$g/ml level was more useful for in vitro fertilization.

• Journal of Microbiology and Biotechnology
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• 제23권9호
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• pp.1327-1338
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• 2013

The humanized anti-hepatocyte growth factor (HGF) monoclonal antibody (mAb) YYB-101 is a promising therapeutic candidate for treating various cancers. In this study, we developed a bioprocess for large-scale production of YYB-101 and evaluated its therapeutic potential for tumor treatment using a xenograft mouse model. By screening diverse chemically defined basal media formulations and by assessing the effects of various feed supplements and feeding schedules on cell growth and antibody production, we established an optimal medium and feeding method to produce 757 mg/l of YYB-101 in flask cultures, representing a 7.5-fold increase in titer compared with that obtained under non-optimized conditions. The optimal dissolved oxygen concentration for antibody production was 70% $pO_2$. A pH shift from 7.2 to 7.0, rather than controlled pH of either 7.0 or 7.2, resulted in productivity improvement in 5 L and 200 L bioreactors, yielding 737 and 830 mg/ml of YYB-101, respectively. The YYB-101 mAb highly purified by affinity chromatography using a Protein A column and two-step ion exchange chromatography effectively neutralized HGF in a cell-based assay and showed potent tumor suppression activity in a mouse xenograft model established with human glioblastoma cells.