• Journal of Embryo Transfer
• /
• v.13 no.2
• /
• pp.117-125
• /
• 1998

본 연구는 Percoll,Swim-up 그리고 Glass-wool 여과법의 세가지 정자분리 방법에 대한 정자 회수율, 생존율과 첨체반응율, 그리고 체외수정 후 배양시간에 따른 전핵형성율,수정란의 발달율과 세포할구수에 대한 효과를 조사하고자 실시하였다. 도축된 암소로 부터 채취한 난자를 22시간 체외배양 후 성숙된 난자를 체회수정시켰다. 수정 후 배양시간에 따라 존핵율을 조사하였으며 48시간에 분할율,192시간에 배반포기 발달율 및 세포할구수를 각각 비교 조사하였다. 정자의 첨체반응과 생존율은 처리군간에 차이가 없으나 회수율에 있어서 percoll 처리군이 다른 두 처리군보다 유의적으로 높았다(p<0.001).수정 후 배양시간별 전핵형성에 있어서도 percoll 처리군이 다른 두 처리군보다 빨리 진행됨을 볼 수 있다. 분할율에 있어서는 처리군간에 유의적 차이가 없으나.배반포기 발달율과 세포할구수에 있어서는 pecoll 처리군이 다른 두 처리군보다 유의적으로 높았다(P<0.05). 이상의 결과로 보아 percoll 처리에 대한 정자분리 방법은 정자 회수율이 높고 수정시 전핵형성 시간이 단축되어, 그 결과로 배반포기 발달율과 수정란의 세포할구수에 효과적임을 알 수 있었다.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.32 no.6
• /
• pp.748-752
• /
• 1999

This study was carried out to compare the growth and fatty acids composition of the rotifer (Brachionus rotundiformis) cultured in high density by the various enrichments and culture methods. The rotifer fed on condensed freshwater Chlorella was enriched with $\omega$-yeast, Algamac, Super Selco and marine Chlorella. In another culture method, the rotifer was cultured with enrichment supplements for 6 hours after feeding with condensed freshwater Chlorella supplement for 18 hour. The rotifer fed with condensed marine Chlorella for 24 hours without fieshwater Chlorella was used as a control group. Culture tanks (5 $\ell$ working volume) was immersed in a water bath ($28^{\circ}C$). The density of rotifer and dissolved oxygen level in water was stable in control group of rotifer cultured with condensed marine Chlorella for 24 hours and the n-3 HUFA content of rotifer was the highest among the rotifer culture methods. However, the density of rotifer and dissolved oxygen level in the groups of rotifers enriched with $\omega$-yeast, Algamac and Super Selco by methods were drastically decreased, The n-3 HUFA contents of rotifers enriched by Super Selco were higher than those of rotifer enriched by either $\omega$-yeast or Algamac in both methods. The results from this experiment indicated that supplementation of condensed marine Chlorella for 24 hour by the semi-continuous culture was effective for the improvement of the nutritional value of rotifer and it could provide the stable growth condition for rotifer culture in high density.

• Development and Reproduction
• /
• v.11 no.3
• /
• pp.205-217
• /
• 2007

As an initial step toward better understanding of the molecular mechanism of estrogen-dependent growth in myoma, an optimal primary cell culture condition has been developed and examined by this study. Myoma and myometrium were cultured by two different methods. Culture stability and $E_2$-responsiveness in stable culture were studied. The culture of digested tissue pieces(Method 2) was found to be a stable culture method for the myoma and myometrium showing a favorable response to estrogen. mRNA expression of PR, IGF-1 and IGF-1 receptor genes was enhanced by $E_2$. The gene responses to $E_2$ were higher in myoma compared with myometrium. Moreover, these responses were more expressive in tissues than in the surrounding cells in primary culture of normal myometrium and myoma, implying a vital role of cell communication through the extracellular matrix in maintaining the estrogen-responsiveness. The development of an improved cell culture system for myoma provides an in vitro tool to further investigate the basis of the tumor formation.

• Journal of Plant Biotechnology
• /
• v.34 no.1
• /
• pp.1-6
• /
• 2007

Effect of BA concentrations and culture methods on in vitro plant multiplication from shoot-tip cultures of Wasabia japonica was studied. Shoot-tips with leaf primordia and apical meristem were cultured on MS basal medium for all the experiments. Liquid medium for 2 weeks followed by semi-solid medium for 4 weeks containing 1.0 mg/L BA was the best to number of shoots (22.8) and shoot length (3.5 cm). Shoots proliferated could be divided into ca. 5 to 11 of cultures for the multiplication of plantlets. Divided plantlets showed root formation (90%) well onto MS basal medium without growth regulators like IBA and NAA. After rooting, all the plantlets transferred into the pots containing composed soil (bio-media Co., peatmoss $8{\sim}10%$, coir dust $66{\sim}70%$, zeolite $13{\sim}17%$, vermiculite $3{\sim}7%$, perlite $2{\sim}4%$) and grown well into whole plants with multiple shoots.

• Proceedings of the KSAR Conference
• /
• 2003.06a
• /
• pp.89-89
• /
• 2003

본 연구는 난소의 황체를 체외배양시 TGF-$\beta$1의 황체내 발현을 조사하기 위해 수행되었다. 돼지 황체는 축산기술연구소에서 사육중인 돼지(체중 145$\pm$kg) 12두로부터 발정을 유도시켜 배란 후 약 48시간째 도축하여 난소를 회수하였다. 회수된 난소로부터 황체를 분리하여 세절한 후 0.25% collagenase용액(0.025mg DNase, 50mM EDTA, 50mM Dithio-threitol)으로 37$^{\circ}C$의 진탕 수조에서 30분간 배양하여 황체세포를 분리 회수하였다. 회수된 황체세포는 D-MEM용액(GIBCO, 10% FCS와 antibiotics 첨가)으로 2회 세척하여 1$\times$$10^{6}$live cell/$m\ell$이 되도록 희석하여 24 well culture plate(Corning, New Tork 14831)에 분주하여 $CO_2$ 배양기($CO_2$: 5%)에서 24시간 간격으로 2회 배양액을 교환해 48시간 동안 배양하였다. 배양된 황체 세포는 immunocytochemistry 방법으로 TGF-$\beta$1의 발현을 관찰함과 동시에 황체조직도 같은 방법을 사용하여 TGF-$\beta$1 의 발현 유ㆍ무을 관찰하였다. 그 결과 황체세포 그리고 황체 조직 뚜렷한 TGF$\beta$1의 발현을 확인할 수 있었다. 이 결과로서 TGF$\beta$1은 황체기능을 유지하는데 하나의 인자로서 작용하며 다른 인자들과의 상호작용을 시사하고 있다.

• Food Industry And Nutrition
• /
• v.8 no.3
• /
• pp.45-51
• /
• 2003

본 연구는 생물공학적인 방법을 도입하여 폐기되는 신선초박에 H. erinaceum의 균사체를 발효시킨 배양생성물을 이용하여 기능성 건강 음료 개발을 검토 하고자 하였다. 1. 종균제조공정개발 : 기본 배지의 선발에서 Hericium erinaceum의 균사 생육에 적합한 배지를 선발하기 위하여 10여종의 고체배지를 사용하여 균사 생육 및 밀도를 조사한 결과는 YMPG 배지에서 59.8mm/14days로 균사 생육이 가장 우수한 것으로 나타났고, 최적 온도는 20-$25^{\circ}C$ 범위에 가장 생육이 좋았으며, 배지의 pH를 조절하여 균사생육을 조사한 결과, pH는 5.5, 접종비는 전배양액 9%(v/v), 배양에 적합한 배지액량은 50mL, 최적교반속도는 120rpm이었다. 이러한 최적 조건 하에서 배양 경시 변화를 살펴 본 결과 당은 거의 일정한 속도로 감소하는 반면에 건조균체량은 배양 8일째까지 증가하다가 더 이상 변화가 없었다. 2. 발효공정개발: 수분함량이 200%(v/v)에서, pH5에서의 생육속도는 90mm/30 days, $25^{\circ}C$에서의 균사생육속도는 89mm/30days로 각각 H. erinaceum균사의 생육이 가장 우수한 결과를 얻었다 3.추출공정 및 시제품 제조: 녹즙을 생산한 후 폐기되는 신선초박에 액체 종균을 접종하여 ,40일 동안 배양시켜 생육 상태가 우수한 배양생성물만을 선별하여, 열수추출방법으로 10$0^{\circ}C$, 10시간 추출한 것을 음료제조의 원료로 하고, 음료의 기호성을 향상시키기 위해 유기산 및 한방추출물을 첨가하는 균사체음료의 조성비를 얻었다.

• Korean Chemical Engineering Research
• /
• v.47 no.2
• /
• pp.220-229
• /
• 2009

Strain improvement and morphology investigation in bioreactor cultures were undertaken in suspended cultures of Phellinus linteus mycelia for mass production of protein-bound polysaccharides(soluble ${\beta}$-D-glucan), a powerful immuno-stimulating agent. Phellineus sp. screened for this research was identified as Phellinus linteues through ITS rDNA sequencing method and blast search, demonstrating 99.7% similarity to other Phellinus linteus strains. Intensive strain improvement program was carried out by obtaining large amounts of protoplasts for the isolation of single cell colonies. Rapid and large screening of high-yielding producers was possible because large numbers of protoplasts ($1{\times}10^5{\sim}10^6\;protoplasts/ml$) formed using the banding filtration method with the cell wall-disrupting enzymes could be regenerated in relatively high regeneration frequency($10^{-2}{\sim}10^{-3}$) in the newly developed regeneration medium. It was demonstrated that the strains showing high performances in the protoplast regeneration and solid growth medium were able to produce 5.8~6.4%(w/w) of ${\beta}$-D-glucan and 13~15 g/L of biomass in stable manners in suspended shake-flask cultures of P. linteus mycelia. In addition, cell mass increase was observed to be the most important in order to enhance ${\beta}$-D-glucan productivity during the course of strain improvement program, since the amount of ${\beta}$-D-glucan extracted from the cell wall of P. linteus mycelia was almost constant on the unit biomass basis. Therefore we fully investigated the fungal cell morphology, generally known as one of the key factors affecting cell growth extent in the bioreactor cultures of mycelial fungal cells. It was found that, in order to obtain as high cell mass as possible in the final production bioreactor cultures, the producing cells should be proliferated in condensed filamentous forms in the growth cultures, and optimum amounts of these filamentous cells should be transferred as active inoculums to the production bioreactor. In this case, ideal morphologies consisting of compacted pellets less than 0.5mm in diameter were successfully induced in the production cultures, resulting in shorter period of lag phase, 1.5 fold higher specific cell growth rate and 3.3 fold increase in the final biomass production as compared to the parallel bioreactor cultures of different morphological forms. It was concluded that not only the high-yielding but also the good morphological characteristics led to the significantly higher biomass production and ${\beta}$-D-glucan productivity in the final production cultures.

• 한국생태학회:학술대회논문집
• /
• 1998.05a
• /
• pp.122.1-122.1
• /
• 1998

• KSBB Journal
• /
• v.15 no.2
• /
• pp.134-138
• /
• 2000

Addition of carbon and nitrogen source to an alcohol distillery wastewater was tried to increase the cell concentration of a b baker's yeast cultivated in that wastewater. Carbon was found to be primary limiting nutrient and nitrogen secondary limiting o one. Glucose addition increased the cell concentration 1.3 times higher than no addition, and both glucose and $(NH_4)_2S0_4$ a addition did 5.8 times. A fed-batch cultivation by the automatic addition of glucose and ammonium was executed. Added g glu$\infty$se was automatically controlled to low concentration by a method using DO as control parameter. Ammonium was a automatically added as NH40H used as pH $\infty$ntrol agent after initiating glucose addition. By this simple cultivation method t the cell concentration $\infty$내d be efficiently increased from 2.6g/L to 12.0g/L, and maximum specific growth rate and biomass y yield to glu$\infty$se were $0.18hr^{-1}$ and about 0.54g/g respectively. By increasing cell concentration, COD of the wastewater m media could be additionally reduced by about 22%.

• Korean Journal of Plant Tissue Culture
• /
• v.24 no.6
• /
• pp.369-374
• /
• 1997

Immature flower buds of 'Desio' carnation were cultured on MS agar medium supplemented with 1 ㎎/L 2,L-D. Embryogenic calli were formed from 5-10% of the buds less than 20 ㎜ in length, but only non-embryogenic calli were produced from explants of shoot apex leaf, internode, and flowere buds larger than 20 ㎜. The same method was applied to 16 cultivars of cut Sower carnation and embryogenic calli were obtained in 7 cultivars. Several embryogenic callus lines were selected and maintained through subcultures over 120 weeks without loss of embryogenic competence. The embryogenic cultures were also proliferated rapidly in liquid agitation cultures using MS medium supplemented with 1mg/L 2,4-D. Numerous embryos were formed on the periphery of the cell aggregates upon transfer to auxin-free MS agar medium. Plantlets were transplanted in potting soil and grown to bloom in six months.