• 한국가축번식학회지
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• 제23권4호
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• pp.393-398
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• 1999

Somatic cell nuclear transfer is an efficient technique for the multiplication of elite livestock, engineering of transgenic animals, cell therapy and xenotransplantation, and analyzing the interactions between nucleus and cytoplasm, for various agricultural, biomedical and research purposes. Since the first somatic cell clone lamb was born, tremendous progress has been made toward developing technology for animal cloning. Viable farm animals and mice have now been produced by nuclear transfer using various fetal and adult somatic cells as nuclei donors. Transgenic clones were also produced from nuclear transfer of transfected somatic cells. In the future, somatic cell nuclear transfer will provide more numerous opportunities, both in basic and appled research as well as immediate uses in the generations of superior clone and transgenic animals. However, further technology refinement and improved understanding of the process are essential for commercial and basic research applications.

• 한국식생활문화학회지
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• 제27권6호
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• pp.743-750
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• 2012

A tissue cultured wild ginseng (TCWG) suspension was inoculated with lactic acid bacteria and fermented to improve the functionality of TCWG. The utilization of TCWG was increased directly using the freeze-dried powder. The optimal ratio of TCWG powder and water for fermentation was 1:19 (5%), which was selected by measuring the fluidity and viable cell count according to concentration. The effects on ADH activation and immune cell activation by each ferments with 10 kinds of Lactobacillus sp. strains were examined. The ferments with the Lactobacillus casei KFRI 692 strain showed 5.4 times higher ADH activity and 1.3 times higher ALDH activity than the non-fermented TCWG powder (control). The level of NO production and cytotoxicity was also measured by Raw 264.7 cells. The ferment with the Lac. casei KFRI 692 strain showed the highest level of NO production and lower cytotoxicity than the others. Therefore, the Lac. casei KFRI 692 strain was selected as a strain for fermentation of a TCWG suspension to maximize its functionality. To identify the optimal fermentation time of the selected Lac. casei KFRI 692 strain on the 5% TCWG suspension, the viable cell count of lactic acid bacterial and the changes in pH were observed for 72 hours. 24-hrs was found to be the optimal fermentation time. In this way, fermented TCWG with lactic acid bacteria showed higher ADH activation efficacy and immune cell activation than non-fermented TCWG.

• Journal of Microbiology and Biotechnology
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• 제12권3호
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• pp.444-448
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• 2002

A $CaCO_3$-alginate beads system was developed for high cell density cultivation of Bifidobacterium longum and the cost-effective media were also screened. In batch process with $CaCO_3$, beads, two strains of B. longum showed both the highest viable cells and optical density in TPY medium, resulting in maximum optical density and viable cell counts of 12.40, $2.22{\times}10^10$ cfu/ml for B. longum ATCC 15707 and 13.71, $3.93{\times}10^10$ cfu/ml for B. longum HLC 3742. Released size distribution, according to $CaCO_3$-alginate bead size preparation, was smaller than others. These results were also examined by observing their morphology. The skim milk-based medium was most adequate to cultivate B. longum as the cheapest medium, and $10\%$ skim milk supplemented with $2\%$ glucose and $1\%$ yeast extract was a suitable medium, supporting the growth to $5.57{\times}10^10$ cfu/ml for ATCC 15707 and $6.82{\times}10^9$ cfu/ml for HLC 3742. During the long-term storage at $4^{\circ}C\;and\;-20{\circ}C$, B. longum cultivated with $CaCO_3$ beads had the highest stability. Consequently, $CaCO_3$-alginate beads buffer was found to be useful not only to cultivate B. longum but also to preserve cultures.

• 한국식품영양과학회지
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• 제28권5호
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• pp.997-1002
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• 1999

The aim of the present study was to investigate effects of food preservative addition and changes in composition of herb extract product on the growth of spoilage yeast, Zygosaccharomyces sp. Herbs such as Panax ginseng, Cinnamomum cassia, Lycium chinense, Zyzyphus juiuba and Jingiber officinale were altogether put into water and essence was extracted at 80oC, and then the extract was concentrated at 75oC. The herb extract product was made by adding vitamins, amino acids and honey to the concentrated herb extract. The amount of gas produced from the herb extract product was increased as inoculated cell number increased but decreased as Brix concentration increased. Gases were produced in small amount when incubation was made at 4oC but large amounts of gases were produced at 25 or 40oC of incubation. The gas production and growth of Zygosaccharomyces sp. were measured after browning reaction was induced by heating at 85oC for 12 hours. It appeared that heating treatment did not induce any significant change in the gas production and growth of the cell. The effects of addition of various sugar to the herb extract produce were also invesigated. Amounts of gas production were in the order of glucose>sucrose>oligosaccharide>stevioside. The viable cell count was measured as 6.0$\times$107 CFU/g when glucose was added to the herb extract product. The viable cell counts were 5.0$\times$106, 3.0$\times$103, and 3.0$\times$102 CFU/g in sucrose, oligosaccharide and stevioside added herb extract product, respectively. The amount of gas production from the herb extract product was remarkably reduced by addition of such food preservatives as sodium benzoate and DF 100. TLC(thin layer chromatography) chromatogram of the herb extract showed stability of the herb extract in the above treatments.

• KSBB Journal
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• 제9권1호
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• pp.48-54
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• 1994

HEK 세포 배양액 중에 존재하는 scu-PA와 tc-PA의 보다 정확한 측정을 위해 fibrin plate법과 기존의 amidolytic 방법을 변형시킨 측정법을 이용했다. 1%의 저혈청 배지를 이용한 T-flask 배양에서 $1.65{\times}10^6$(viable cells/ml)의 최대 세포수에 도달하여 1670(IU/ml)의 scu-PA 생산량을 보였으며 평균 10% 미만이 변환율을 보였다. 또한 Spinner vessel에서의 회분배양시 최대 세포수가 $4.43{\times}10^6(total cells/ml)$ 와 1560(IU/ml)의 scu-PA 생산량을 나타냈으며 평균 11.4%의 변환율을 보였다. 연속배양에서는 0.449(1/day)의 비생육속도와 $3.13{\times}10^{-4}(IU/cell)$의 비생산속도를 보였으며 평균 10.18% 정도의 전환율을 보였다. 이는 회분식 및 유가식 배양 결과와 큰 차이가 없으며 배양공정에 관계없이 약 90% 이상의 회수율이 가능하다는 것을 의미한다.

• Journal of Microbiology and Biotechnology
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• 제20권11호
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• pp.1534-1538
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• 2010

Fed-batch cultures of Hansenula polymorpha were studied to develop an efficient biosystem to produce recombinant human serum albumin (HSA). To comply with this purpose, we used a high-purity oxygen-supplying strategy to increase the viable cell density in a bioreactor and enhance the production of target protein. A mutant strain, H. polymorpha GOT7, was utilized in this study as a host strain in both 5-l and 30-l scale fermentors. To supply high-purity oxygen into a bioreactor, nearly 100% high-purity oxygen from a commercial bomb or higher than 93% oxygen available in situ from a pressure swing adsorption (PSA) oxygen generator was employed. Under the optimal fermentation of H. polymorpha with highpurity oxygen, the final cell densities and produced HSA concentrations were 24.6 g/l and 5.1 g/l in the 5-l fermentor, and 24.8 g/l and 4.5 g/l in the 30-l fermentor, respectively. These were about 2-10 times higher than those obtained in air-based fed-batch fermentations. The discrepancies between the 5-l and 30-l fermentors with air supply were presumably due to the higher contribution of surface aeration over submerged aeration in the 5-l fermentor. This study, therefore, proved the positive effect of high-purity oxygen in enhancing viable cell density as well as target recombinant protein production in microbial fermentations.

• 한국식품과학회지
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• 제33권4호
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• pp.444-450
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• 2001

고추장에 초고압-열 병합처리법을 적용하여 미생물 살균효과 및 품질변화와 $37^{\circ}C$ 저장 중 품질변화를 측정하였다. 고추장을 680 MPa/30분에서 $49{\sim}73^{\circ}C$로 처리하였을 때 생균수는 $0{\sim}3$ log cycle 감소하였고. $73^{\circ}C/30$분에서 $380{\sim}680\;MPa$로 처리하였을 때 $0{\sim}3$ log cycle 감소하였고, $73^{\circ}C/680\;MPa$에서 $10{\sim}70$분 처리하였을 때 $2{\sim}5$ log cycle 감소하였다. 고추장의 pH, 적정산도, 아미노태질소 함량, 환원당 함량, 에탄올 함량 등 이화학적 성질은 초고압-열 병합처리 여부와 처리 조건에 관계없이 유의적인 차이가 없었다. 초고압-열 병합처리한 고추장의 L, a, b값은 무처리 고추장에 비하여 유의적으로 높았다. 고추장을 $37^{\circ}C$에서 저장 중 생균수와 품질 변화를 측정하였다. 고추장의 생균수는 저장기간에 따라 감소하는 경향을 보였다. 생균수는 380 Mpa/30분 처리군이 처리직 후 $1.88{\times}10^6$에서 저장 120일 후 $1.94{\times}10^4$로 약 2 log cycle 감소한 반면, 680 MPa/70분 처리군은 저장 초기 $4.00{\times}10^1$에서 60일 후 검출되지 않았다. 저장기간에 따라 고추장의 pH는 유의적으로 감소하였고, 적정산도는 증가하였다. 아미노태질소 함량은 저장기간에 따라 유의적으로 감소하였고, 환원당 함량은 다소 증감을 반복하였으며, 에탄올 함량은 저장기간에 따라 간헐적으로 다소 감소하는 경향을 보였다. 고추장의 L, a, b값은 저장기간에 따라 유의적으로 큰 폭으로 감소하였다. 전체적으로 380 MPa/30분 처리군보다 680 MPa/70분 처리군의 변화폭이 더 컸다.

• 한국식품위생안전성학회지
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• 제13권2호
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• pp.77-82
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• 1998

생약제 농축액의 안정성 확보하기 위한 기초자료를 제공하고자 생약제 농축액에서 수분 활성도 변화에 따른 미생물 성장 변화 및 일부 이화학적 특성 변화를 조사하였다. 수분 활성도를 각각 0.86, 0.89, 0.69로 조정한 생약제 농축액을 $40^{\circ}C$에서 180일 동안 저장한 후 세균수의 생균수를 측정한 결과 수분활성도 0.86(수분 활성도 63.85%) 시료의 경우 초기 생균수가 18/g에서 90일 저장시 80/g으로 180일 저장시 190/g으로 증가하였다. 수분활성도 0.80(수분함량 43.04%) 경우에는 초기생균수가 24/g에서 90일 저장시 83/g으로, 180일 저장시 170/g으로 증가하였다. 그러나 수분활성도 0.69(수분함량 33.05%) 경우에는 초기 생균수가 16/g에서 90일 및 180일 저장시 각각 2-/g, 25/g으로 큰 균수가 증가를 나타내지 않았다. 한편 수분 활성도에 따른 병원성 미생물의 성장은 수분활성도가 낮을수록 저해되었으며 C. albicans, A. niger의 경우 $28^{\circ}C$에서 30일 저장 후 초기 생균수가 150/g, 140/g에서 각각 30/g, 20/g으로 크게 감소하였고, E. coli, S. aureus, P. aeruginosa의 경우에도 수분활성도 0.69 시료에서는 $37^{\circ}C$에서 30일 저장 후 생균수가 0로 나타나서 균은 완전히 사멸되었다. 수분활성도 0.86, 0.80, 0.69 시료에서 pH의 변화는 180일 저장시의 pH에서 초기 pH를 뺀 pH의 감소 폭이 각각 0.84, 0.72, 0.31로 나타나서 수분활성도가 낮을수록 pH의 감소폭이 적은 경향이었다. 생약제 농축액에 포함된 인삼 중의 지표성분인 사포닌은 180일 저장한 시료에서 측정한 사포닌함량에서 저장 초기 시료의 함량을 뺀 사포닌 함량의 감소폭은 0.12~0.30%로 나타나서 비교적 작은 편이었다. 또한 생약제 농축액에 감미료, 산미료 등의 첨가물을 첨가하여 $40^{\circ}C$에서 60일 및 180일 동안 저장한 후 TLC로 사포닌을 조사한 결과 모든 사포닌이 확인되어 이러한 조건에서 사포닌은 안정한 것으로 생각된다.

• KSBB Journal
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• 제4권2호
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• pp.143-149
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• 1989

하이브리도마 세포를 고농도 배양하기 위하여 세포 침전장치를 고안하여 사용하였다. 위로 올라갈수록 직경이 넓어지는 침전조는 세포의 침전이 잘 일어나지만 침전된 세포가 침전조 벽에 누적되는 결점을 보였고, 아래 위의 직경이 균일한 침전조는 희석율이 높아질수록 유출되는 세포가 증가하는 문제점을 나타내었다. 특히 유출되는 세포속에는 dead cell 보다 viable cell의 비율이 높아 배양기 내의 dead cell이 급격히 농축되는 현상을 보였다. 그러나 침전조를 적절히 고안하여 세포 누적 현상을 막고 동시에 세포 유출을 완화시킬 경우 세포농도 $5{\times}10^6$ cells/ ml에서 일주일간 배양이 가능했다. cell viability의 측정이 용이하므로 침전조를 이용한 하이브리도마 고농도 배양의 특성을 연구하는데 많은 도움을 줄 수 있을 것으로 판단된다.

• Journal of Periodontal and Implant Science
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• 제31권3호
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• pp.597-610
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• 2001

Normal gingival fibroblasts functioning is fundamental for the maintenance of periodontal connective tissue as well as wound healing. Nicotine have been found to affect DNA synthesis and cell proliferation, which appear to depend on the type of cells. This in vitro study was done to determine the effects of nicotine, a major component of tobacco, on cell proliferation, viability, activity, cell cycle distribution, and expression of cell cycle regulatory proteins in human gingival fibroblasts. Nicotine has been tested for 2 days or 4 days in 5 different concentrations; $0.1{\mu}g/ml$; $1{\mu}g/ml$; $10{\mu}g/ml$; $100{\mu}g/ml$; $1000{\mu}g/ml$. To assess cell proliferation and viability, viable and non-viable cells were counted by hemocytometer; to evaluate cellular activity, MTT assay was employed; to analyze cell cycle distribution, fluorescent propidium iodide-DNA complex were measured using fluorocytometer; to determine the expression of cell cycle regulatory proteins, western blot analysis was performed. After 2 days and 4 days incubation respectively, at concentrations of $1{\mu}g/ml$ - $1000{\mu}g/ml$, nicotine significantly inhibited proliferation comparing to non-supplemented controls. The cell viability was significantly decreased after 2 days and 4 days at concentrations of $1{\mu}g/ml$ - $1000{\mu}g/ml$ and at $10{\mu}g/ml$ - $1000{\mu}g/ml$ respectively. After 2 days and 4 days, the cellular activity was significantly decreased at concentrations of $10{\mu}g/ml$ - $1000{\mu}g/ml$. Treatment with $100{\mu}g/ml$ nicotine for 48 hours caused an increase in the proportion of G1-phase cells (from 46.41% to 53.46%) and a decrease in the proportion of S-phase cells (from 17.80% to 14.27%). The levels of cyclin $D_1$ and CDK 4 proteins in nicotine-treated fibroblasts were lower than that of controls, whereas the levels of p16 and pRB were higher than that of controls. These results suggest that the decrease of cell proliferation and lengthened Gap phases (G1) by nicotine may due to the increased expression of p16 and pRB as well as decreased expression of cyclin $D_1$ and CDK 4 in human gingival fibroblasts.