• 제목/요약/키워드: drop-culture

검색결과 119건 처리시간 0.028초

Investigation on the Microbiological and Biochemical Properties of Kimchi in the Solid-state Model System Designed for Fermented Sausages

  • Lee, Joo-Yeon
    • 한국축산식품학회지
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    • 제30권2호
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    • pp.236-242
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    • 2010
  • The objective of this study was to investigate the potential of the application of kimchi LAB as starter culture in the production of fermented sausages. For this, the solid-state model media composed to simulate the substantial conditions of meat mixtures were fermented for 120 h after the treatment with different concentrations of kimchi (0.5, 1.0, 1.5, 3.0, and 5.0%) and lyophilized kimchi-powder (0.2 % and 0.5%). During the fermentation period, the growth of total viable cells and LAB, and the changes of pH and titratable acidity were investigated. The initial LAB counts ranged from 7.18 to 8.34 Log CFU/ mL for kimchi media and from 6.93 to 6.94 Log CFU/mL for kimchi-powder media depending on the added concentrations. The kimchi LAB in this study were not influenced by the immobilized condition for their adaptation and growth by showing no lag phase and thus acted similar as in the submerged medium. The initially increased counts reached around 9 Log CFU/ mL in 12 h independent of the concentrations of a ded kimchi. However, the growth and metabolic activity of kimchi-powder LAB were influenced by the immobilized condition. Supposedly, as the nutrient supply in solid-state depended solely on diffusion, these differences in the souring properties were caused by the LAB topography in the medium matrix. Nevertheless, the differences in the numbers of LAB between two media were less than 0.5 Log units and the pH drop in the solidstate batches was quite rapid and reached low values. Therefore, it can be assumed that kimchi and kimchi-powder LAB showed the utility as the substitute of commercial starter culture even without a rehydrating pretreatment.

Addition of interleukin-6 to mouse embryo culture increases blastocyst cell number and influences the inner cell mass to trophectoderm ratio

  • Kelley, Rebecca L;Gardner, David K
    • Clinical and Experimental Reproductive Medicine
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    • 제44권3호
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    • pp.119-125
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    • 2017
  • Objective: In vitro culture of preimplantation embryos is improved by grouping embryos together in a drop of media. Individually cultured embryos are deprived of paracrine factors; with this in mind, we investigated whether the addition of a single embryo-secreted factor, interleukin-6 (IL-6), could improve the development of individually cultured embryos. Methods: Mouse embryos were cultured individually in $2{\mu}L$ of G1/G2 media in 5% oxygen and supplemented with a range of doses of recombinant mouse or human IL-6. Results: Mouse IL-6 increased hatching at doses of 0.01 and 10 ng/mL compared to the control (93% and 93% vs. 78%, p< 0.05) and increased the total number of cells at a dose of 0.1 ng/mL compared to the control ($101.95{\pm}3.36$ vs. $91.31{\pm}3.33$, p< 0.05). In contrast, the highest dose of 100 ng/mL reduced the total number of cells ($79.86{\pm}3.29$, p< 0.05). Supplementation with human IL-6 had a different effect, with no change in hatching or total cell numbers, but an increase in the percentage of inner cell mass per embryo at doses of 0.1, 1, and 100 ng/mL compared to the control ($22.9%{\pm}1.1%$, $23.3%{\pm}1.1%$, and $23.1%{\pm}1.1%$ vs. $19.5%{\pm}1.0%$, p< 0.05). Conclusion: These data show that IL-6 improved mouse embryo development when cultured individually in complex media; however, an excess of IL-6 may be detrimental. Additionally, these data indicate that there is some cross-species benefit of human IL-6 for mouse embryos, but possibly through a different mechanism than for mouse IL-6.

인간 배아줄기세포의 생식세포로의 분화 및 효소에 의해 분리된 단일줄기세포 배양조건 (Differentiation of Human Embryonic Stem Cells into Germ Cell and Culture Condition for Single Embryonic Stem Cells Dissociated by Enzyme)

  • 지희준;최순영;정다연
    • Clinical and Experimental Reproductive Medicine
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    • 제37권1호
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    • pp.13-23
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    • 2010
  • 목 적: 본 연구는 인간 배아줄기세포를 생식세포로의 분화를 유도하고 효소에 의해 분리된 단일 배아줄기세포의 배양조건을 확립하기 위해 수행하였다. 연구방법: Embryonic body (EB)는 배아줄기세포 (hESCs) colony를 떼어내어 3일간 hanging drop culture 방법으로 작성하였고, 이러한 EB를 retionic acid (RA)와 bone morphogenetic protein-4 (BMP4)를 단독 또는 함께 배양액에 첨가하여 14일간 배양함으로써 생식세포로의 분화를 유도하였다. 분화를 유도한 EB는 생식세포 발현유전자인 c-kit과 VASA의 표지인자를 이용한 면역조직형광법으로 분화여부를 조사하였다. 줄기세포는 Collagenase, Tryple 그리고 Accutase 등의 효소로 각각 분리하였고 분리된 단일세포들의 colony formation rate를 조사하였다. 한편 Rho-associated kinase inhibitor (Y-27632)를 단일세포 배양액에 첨가하여 단일세포 분리과정 중에 발생하는 apoptotic damage를 감소시키고자 하였다. 결 과: Tryple 또는 Accutase를 이용하여 분리한 단일세포가 Collagenase에 의해 분리된 세포에 비해 높은 colony formation rate를 나타내었다. 단일세포를 $5{\times}10^3$ cells/well (4 well dish) 농도로 지지세포 위에 seeding하였을 때 다른 농도의 세포를 seeding한 것에 비해 높은 colony formation rate를 확보하는데 효과적이었다. Y27632의 첨가는 단일세포의 colony formation rate를 유의하게 향상시켰으며 특히 Tryple로 분리한 단일세포에 보다 효과적이었다. EB의 분화유도후 c-kit과 VASA의 표지인자를 이용한 면역조직형광염색은 대조군인 정소조직에 비해 약한 형광염색을 나타내었다. 결 론: Tryple을 이용한 단일세포 분리가 건강한 단일세포를 얻는데 가장 효율적이었으며 Y27632 의 첨가는 단일 세포의 생존 및 colony formation에 유익하다는 것을 확인하였다. 다른 연구와는 달리 본 연구에서는 단지 생식세포 표지인자의 희미한 형광염색만을 관찰하였는데 이러한 결과는 본 연구의 분화유도기간이 상대적으로 짧았던 것이 원인이었을 것으로 생각된다.

Lactobacillus plantarum (KACC 92189) as a Potential Probiotic Starter Culture for Quality Improvement of Fermented Sausages

  • Ba, Hoa Van;Seo, Hyun-Woo;Seong, Pil-Nam;Kang, Sun-Moon;Kim, Yoon-Seok;Cho, Soo-Hyun;Park, Beom-Young;Ham, Jun-Sang;Kim, Jin-Hyoung
    • 한국축산식품학회지
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    • 제38권1호
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    • pp.189-202
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    • 2018
  • This study was conducted to evaluate the effects of fermenting temperature on the applicability of Lactobacillus plantarum for production of fermented sausages as starter cultures, and its applicable efficiency was also compared with those inoculated with commercial starter culture or non-inoculated control. The L. plantarum isolated from a naturally-fermented meat, identified by 16S rDNA sequencing and again identified by de novo Assembly Analysis method was used as a starter culture. Six treatments: 3 with L. plantarum at different fermenting temperatures (20, 25 and $30^{\circ}C$), and other 3 treatments (1 with commercial starter culture, 1 with its mixture with L. plantarum and 1 non-inoculated control) fermented under the same conditions ($25^{\circ}C$) were prepared. Results revealed that the fermenting temperature considerably affected the pH change in samples added with L. plantarum; the highest pH drop rate (1.57 unit) was obtained on the samples fermented at $30^{\circ}C$, followed by those at $25^{\circ}C$ (1.3 unit) and $20^{\circ}C$ (0.99 unit) after 4 days fermentation. Increasing the temperature up to $30^{\circ}C$ resulted in significantly lower spoilage bacteria count (5.15 log CFU/g) and lipid oxidation level in the products inoculated with L. plantarum. The sensory analysis also showed that the samples added with L. plantarum at $30^{\circ}C$ had significantly higher odor, taste and acceptability scores than those fermented at lower temperatures. Under the same processing condition, although the L. plantarum showed slightly lower acidification than the commercial starter culture, however, it significantly improved the eating quality of the product.

NCSU-23과 PZM 배양액내 첨가된 Macromolcule이 돼지 체외수정란의 발육에 미치는 영향 (Effect of Macromolecules Supplemented to NCSU-23 and PZM Culture Media on in vitro Development of Porcine Embryos)

  • 김수;이소현;김대영;강성근;이병천;황우석
    • 한국수정란이식학회지
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    • 제18권1호
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    • pp.35-41
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    • 2003
  • 본 연구에서 배양액 내 첨가된 macromolecule이 돼지 체외수정란의 체외 발달율에 미치는 영향과 배양소적 내 첨가된 FBS가 후기배발달에 미치는 영향에 대하여 알아보았다. 이에 대한 결과를 요약 하면 다음과 같다. 1. NCSU-23을 기본 배양액으로 사용시 BSA (4mg/ml), FBS (10%), PVA (3 mg/ml)가 각각 첨가되었을 경우 배발달에 큰 차이를 보이지 않았다. 2. PZM을 기본 배양액으로 사용시 BSA (4mg/ml)가 첨가하였을 경우 2-세포 수정란 발달율과 배반포 발달율이 가장 높았으며, 이것은 NCSU-23에 비하여 PZM 배양액에는 후기배의 에너지원으로 사용하는 글루코스가 포함되어 있지 않기 때문으로 사료 된다. 본 연구를 통하여 돼지 수정란의 체외배양에 사용되는 배양액은 종류와 조성에 따라 적절한 macromolecule이 첨가되어 초기배와 후기배의 발달을 모두 촉진시킬 수 있어야 함을 알 수 있었다.

20대 남성 체형 특성에 따른 토르소 원형 개발 연구 (Development of Torso Pattern according to the Physical Types of Men in 20s)

  • 황은경;김인숙
    • 복식문화연구
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    • 제12권3호
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    • pp.415-428
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    • 2004
  • The purpose of this thesis was to develop torso patterns according to the physical types of men in 20s. The procedure and results are as follows; 1. Of the men in 20s meeting the national average of physique, 4 men were selected and classified into 2 physical types according to their chest and waist circumference drops. One group had 20㎝ drop (Y type) while the other had 16㎝ drop (N type). Through evaluation performed by clothing construction professionals on the 4 upper bodice blocks drafted according to the existent drafting method and dressed on the 4 men, several problems have been found. These problems were adjusted and supplemented to make two new blocks. The fit of the new blocks were evaluated and proved to be satisfactory. 2. The following is the adjustments made to the existent men's torso patterns which had been utilized as the objects the first evaluation experiment. ① Though the back waist length of the pattern from the existent drafting method covered the center back length of the body in both Y type and N type, the front length did not causing it to stand away from the body. To adjust this, 2.0㎝ has been added to the center front length of each pattern so that the waist line could make a bar level to the ground. ② The shoulder line of the pattern from the existent drafting method had the tendency to fall backward. To make the shoulder line to fall in place, it has been moved 1.0㎝ to the front. 0.5㎝ has been added to the should length. ③ The neckline had a tendency to climb up. It has been lowered by 0.5㎝ until the line touched the center front neck point. ④ Though different in degree, the neck circumference did not allow enough width for both physique type causing the neckline to pull at side neck point with diagonal crease. To adjust this, 0.3㎝ and 0.6㎝ has been added to the Y type and N type respectively so that the neckline would touch the side neck point and the neckline could naturally fall into its original position. ⑤ Though different in degree, there was not enough space at the armhole causing wrinkles around this area. Therefore, 0.25㎝ and 0.5㎝ has been added to the front and back of the armholes of the Y and N types respectively. The armhole was made 1.0㎝ deeper only for the N type. ⑥ 1.0㎝ in the front and 0.5㎝ in the back were added to the side scam for the Y type while 0.5㎝ in the front and 0.25㎝ in the back were added for N type. This eliminated the unwanted wrinkles to give the silhouette a smooth look.

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한우에 있어서 난포란의 체외성숙에 영향을 미치는 요인에 관한 연구 (Study on Factors affecting in vitro Maturation of Follicular Oocytes in Korean Native Cattle)

  • 서태광;정범식;김규현;김익수;류재웅;박수봉;박항균
    • 한국가축번식학회지
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    • 제14권4호
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    • pp.263-271
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    • 1990
  • This study was carried out to investigate the factors affecting maturation in vitro of follicular oocytes in Korean Native Cattle. The bovine ovaries were obtained at a slaughter house and the follicular oocytes were recovered by aspirating the follicular fluid from the visible follicles of 3~6mm. The bovine oocytes were matured in vitro in TCM-199 containing FCS and hormones. The effects of TCM-199 salt type, number of oocytes per drop, incubation time and co-culture with granulosa cells on maturation of oocytes, were investigated. The results obtained are summarized as follows. 1. The maturation rates of follicular oocytes cultured for 22, 25 and 28 hours in Hank's TCM-199 or Earle's TCM-199 were 59.3, 59.6, 80% and 80.0, 90.0, 93.7%, respectively. The maturation rate of follicular oocytes in Earle's TCM-199 was faster and higher than in Hank's TCM-199(P<0.05). 2. The maturation rates of oocytes were 54.5, 62.5 and 62.0% when cultured the oocytes number 10, 20 and 40 per 200${mu}ell$ drop for 18 hours. 3. The maturation of follicular oocytes in the Korean Native Cattle was induced at 14 hours culture, by giving the maturation rate of 90.0% after 20 hours. 4. The maturation rates were 63.3% and 66.7%, respectigely when the oocytes were co-cultured with granulosa cells from medium-size follicles used immediately after recovery in the presence or absence of hormones added(0.02AU/ml FSH, 10$\mu\textrm{g}$/ml LH and 1$\mu\textrm{g}$/ml estradiol 17-$\beta$). When the oocytes were co-cultured with granulosa cells from medium-size follicles cultured for 3 days, the maturation rates were 20.8% and 76.2%, respectively(P<0.05). 5. The maturation rate were 88.0% and 70.0%, respectively when the oocytes were co-cultured with granulosa cells from large-size follicles used immediately after recovery in the presence or absence of hormones added(0.02AU/ml FSH, 10$\mu\textrm{g}$/ml LH and 1$\mu\textrm{g}$/ml estradiol 17-$\beta$)(P<0.05). When the hormones added(0.02AU/ml FSH, 10$\mu\textrm{g}$/ml LH and 1$\mu\textrm{g}$/ml estradiol 17-$\beta$)(P<0.05). When the oocytes were co-cultred with granulosa cells from large-size follicles cultured for 3 days, the maturation rates were 41.2% and 73.3%, respectively(P<0.05).

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아미노산과 FBS의 첨가가 한우난자의 체외발달에 미치는 영향 (Effects of the Addition of Amino Acids and FBS on the In Vitro Development of Korean Native Cow IVP Embryos)

  • 박흠대;박향;이상진;김재명
    • 한국수정란이식학회지
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    • 제17권3호
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    • pp.211-218
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    • 2002
  • 본 연구는 체외에서 소 난포란유래의 배반포 생산에 있어서 배지 내에 첨가하는 외인성 고정질소 원으로써 아미노산과 FBS의 첨가효과를 검토하였다. 소 난포란의 체외성숙은 TCM-l99용액, 체외 수정은 Fer-TALP용액으로 행하였으며, 체외수정후 24시간째(day 1)의 수정난자를 체외배양에 제공하였다. 체외배양용 기초배지는 YS용액, 기초 배양법은 25개 난자/10 ${\mu}\ell$ 배지의 단순.미소적배양법을 이용하였다. 본 연구의 결과를 요약하면 다음과 같다. 1. 체외 수정란의 배양에 있어서 비필수 아미노산(MEM 유래) 첨가가 무첨가군보다 높은 배반포 발달율을 나타냈다. 2. 체외 수정란의 배양에 있어서 필수 아미노산(RPMI 1640 유래) 첨가가 무첨가군보다 유의하게 높은 배반포 발달율을 나타냈다. (P<0.05) 3. Day 1에 비 필수 아미노산, day 5에 필수 아미노산을 첨가하였을 경우 부화 배반포로의 발생율 및 배반포로의 부화율을 향상시켰다. 4. Day 1에 비필수.필수 아미노산을 첨가한 후 day 3, day 4, day 5에 각각 필수 아미노산만을 배지로부터 제거 시 배반포의 부화율은 현저하게 낮았다. 5. FBS의 첨가시기는 배양 후기(day 5)에 첨가할 수록 배반포율 또는 부화 배반포율이 유의하게 상승하였다(p<0.05). 이상의 결과에서 소 난포란 유래 배반포의 체외생산에 있어서 배지에 첨가하는 외인성 고정 질소 원들의 첨가에 있어서 첨가시기 및 농도를 조절함으로써 양질의 배반포 생산을 향상시킬 수 있는 것으로 사료된다.

미생물비료 생물자원인 불용성인산 가용화 세균의 분리, 동정 및 생리적 특성 (Isolation, Identification and Physiological Characteristics of Biofertilizer Resources, Insoluble Phosphate-Solubilizing Bacteria)

  • 손홍주
    • 미생물학회지
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    • 제39권1호
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    • pp.51-55
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    • 2003
  • 환경친화적인 미생물인산비료를 개발하기 위하여 불용성 인산을 가용화 시킬 수 있는 세균을 콩의 근권 토양으로부터 분리하였다. 분리균주의 분류학적 위치를 검토한 결과, Pantoea agglomerans로 동정되었다. 불용성 인산인 $Ca_{3}(PO_{4})_{2}$)로부터 가용성 인산을 생성하기 위한 최적 배지 및 배양조건은 glucose 3%, $NH_{4}NO_{3}$ 0.1%, $MgSO_{4}\cdot\7H_{2}O$ 0.02%, $CaCl_{2}\cdot\2H_{2}O$, 0.06%,초기 pH 7.5, 배양온도 $30^{\circ}C$이었으며, 최적조건에서 배양 5 일 후,898 mg/L의 인산이 생성되었다. 불용성 인산 가용화 기작은 유기산 생성에 의한 배양액의 pH감소와 밀접한 관계가 있었다. 분리균주는 $CaHPO_{4}$, hydroxyapatite로부터 각각 698,912 mg/L의 가용성 인산을 생성하였으나 $FePO_{4}\cdot\4H_{2}O$, $AIPO_{4}$로부터는 각각 28,19 mg/L.의 가용성 인산을 생성하였다.

Development of Vitrified Bovine Oocytes following Intracytoplasmic Sperm Injection (ICSI)

  • Yeo, H-J;Ock, S-A;Yea, E-H;Lee, H-J;Choe, S-Y;Park, G-J
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2001년도 춘계학술발표대회
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    • pp.6-6
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    • 2001
  • Oocyte freezing has become a prevalent source for related reproductive technologies. This study was carried out to evaluate viability of post-thawed bovine oocyte injected DTT-treated sperm following by two different activation stimuli (Group 1, 5 M ionomycin, 5 min + CR1aa, 3 h . 1.9 mM dimetylaminopurine (DMP), 3 h; group 2, ionomycin + 10 $\mu\textrm{g}$/$m\ell$ cycloheximide(CHX), 5h). The techniques of ultra-rapid freezing used in this study were essentially similar to those of described by Vajta et al (Theriogenology 1999; 52:939-948), Denuded oocytes at 22 h of culture were exposed to cryoprotectant (3.2 M Ethylene glycol, 2.36 M DMSO, 0.6 M sucrose), and followed by freezing in electron microscopic grid. After thawing the oocytes were transferred back into the drop of maturation medium and cultured for additional 2 h before being subjected to ICSI. All eggs were then cultured in CRlaa medium, and transferred into M199+10% FCS on day 4. The culture was maintained until day 9. In Experiment 1, frozen-ICSI eggs were compared on development into blastocyst to those of unfrozen and IVF control. Those eggs were activated with the method of group 2. A higher proportion of unfrozen-ICSI and IVF eggs developed into cleavage and blastocysts than of frozen-ICSI eggs (65% and 13%; 71% and 23% vs. 39% and 8%; P<0.05). In Experiment 2, development and ploidy of embryos made from group 1 were compared to those from group 2. Between groups there did not differ on the rates of development, however, chromosomal abnormality in group 1 was significantly higher than in group 2 (49% vs. 30%; P<0.05). The present result suggests that frozen bovine oocytes can be used for ICSI.

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