연조직염 환자에서 원인균 검출률 향상을 위한 새로운 액체 증균 배지

김현대 ( Hyun Dai Kim ),김신무 ( Shin Moo Kim ),박석돈 ( Seok Don Park ) 대한피부과학회 2008 대한피부과학회지 Vol.46 No.3

Background: Staphylococcus aureus and Group A beta-hemolytic Streptococci are the etiologic agents most commonly associated with cellulitis, but many other bacteria have also been shown to cause this condition. The positive bacterial culture rate is the most important factor in the treatment of cellulitis. However, the positive bacterial culture rate in the commonly used media, tends to be quite low. Objective: The principal objective of this study was to improve the positive culture rate in cellulitis patients by using a new enriched broth. Methods: Brewer modified thioglycollate medium (BTM) and Columbia broth (CB), both of which are widely utilized in clinical bacteriology for enriched growth, were compared with several novel enriched broths. These new enriched broths were mixtures of BTM-CB broth and added growth supplement factors. They included BTM-CB (BC), Modified BTM-CB (MBC) and supplement VX-BTM-CB (VXBC). MBC media included several growth supplements, such as hemin, vitamin K1, VX supplement, and Campylobacter growth supplement. Strains utilized in this study were common pathogens (Staphylococcus aureus, Streptococcus pyogenes, et al.), anaerobes, fastidious pathogens (Bacteroides fragilis, Campylobacter jejuni, Prevotella melaninogenica), uncommon pathogens (Actinobacter baumannii, Enterococcus faecalis, Streptococcus agalactiae). Positive culture rates were evaluated in each medium and measured via spectrophotometry at 660 nm. Results: In vitro, all strains used in this study grew more quickly and densely in MBC media. Conclusion: Our results suggest that MBC media in a new enriched broth may improve bacterial culture rates in cellulitis patients. It will be necessary to study the efficacy of the MBC media in the culturing etiologic agents from tissues of cellulitis patients. (Korean J Dermatol 2008;46(3):310∼318)

Effect of Low Adapted Temperature and Medium Composition on Growth and Erythropoietin (EPO) Production by Chinese Hamster Ovary Cells

Na Young Kim,김정회,Hong Jin Kim 대한약학회 2005 Archives of Pharmacal Research Vol.28 No.2

Temperature and medium composition were changed with the aim of increasing growth and erythropoietin (EPO) production in EPO-producing Chinese hamster ovary (CHO) cells. We used the CHO cell line, IBE, and its derivative, CO5, which over-expresses the first two enzymes of the urea cycle, carbamoyl phosphate synthetase I (CPS I) and ornithine transcarbamoylase (OTC). When supplements were added to the medium at 33 oC, the growth of IBE and CO5 cells increased by 27% and 26%, respectively and the maximum yield of EPO was increased by 40% in both cell lines. The absolute EPO concentration in the CO5 cells was always 55~60% higher than in the IBE cells. In addition, when the two cell lines were continuously cultured with supplements at 33 oC until their growth rates approached those at 37 oC, the growth rates of both IBE and CO5 cells increased by 54% and their maximum EPO levels increased by up to 73% and 56%, respectively. Therefore, the growth and EPO expression levels of CO5 cells increased 2.2-fold and 2.6-fold, respectively, compared to those of the IBE cells. These results indicate that adaptation to lower temperature as well as medium supplementation could be important for improving cell growth and EPO production.

Effect of Low Adapted Temperature and Medium Composition on Growth and Erythropoietin (EPO) Production by Chinese Hamster Ovary Cells

Kim Na Young,Kim Jung Hoe,Kim Hong Jin The Pharmaceutical Society of Korea 2005 Archives of Pharmacal Research Vol.28 No.2

Temperature and medium composition were changed with the aim of increasing growth and erythropoietin (EPO) production in EPO-producing Chinese hamster ovary (CHO) cells. We used the CHO cell line, IBE, and its derivative, CO5, which over-expresses the first two enzymes of the urea cycle, carbamoyl phosphate synthetase I (CPS I) and ornithine transcar-bamoylase (OTC). When supplements were added to the medium at $33\;^{\circ}C$, the growth of IBE and CO5 cells increased by $27\%\;and;26\%$, respectively and the maximum yield of EPO was increased by $40\%$ in both cell lines. The absolute EPO concentration in the CO5 cells was always $55{\sim}60\%$ higher than in the IBE cells. In addition, when the two cell lines were continuously cultured with supplements at $33\;^{\circ}C$ until their growth rates approached those at $37\;^{\circ}C$, the growth rates of both IBE and CO5 cells increased by $54\%$ and their maximum EPO levels increased by up to $73\%\;and\;56\%$, respectively. Therefore, the growth and EPO expression levels of CO5 cells increased 2.2-fold and 2.6-fold, respectively, compared to those of the IBE cells. These results indicate that adaptation to lower temperature as well as medium supplementation could be important for improving cell growth and EPO production.

Factors Affecting Primary Cultures of Abalone Haliotis discus hannai Ovary-dissociated Cells and General Culture Aspects

Ryu, Jun Hyung,Nam, Yoon Kwon,Gong, Seung Pyo The Korean Society of Fisheries and Aquatic Scienc 2015 Fisheries and Aquatic Sciences Vol.18 No.1

We investigated factors affecting primary cultures of Pacific abalone Haliotis discus hannai ovary-dissociated cells to identify general aspects of their early-phase culture. Ninety-seven cell populations derived from 30 individuals were cultured in different media with varying compositions of medium supplements, and initial attachment, subculture, and survival for ${\geq}10$ weeks were assessed according to medium composition and individual. We also examined the time required for subculture and the rate of cell death according to both culturing period and passage number within 10 weeks. A lack of fetal bovine serum (FBS) and hemolymph significantly inhibited the growth of cultured cells, while we detected no significant effect of medium composition on initial cell attachment. Through data reallocation, with the omission of data from cell populations cultured in FBS-free and hemolymph-free media, we showed that growth inhibition was also affected by individual differences among the abalones used. During the culture, we observed four different types of cell morphology. Moreover, considerable time was required for subculture-18.4 and 19.5 days for first and second subcultures, respectively-and cell death did not occur within 30 days or for passage 0. Our results will provide valuable information for developing universal cell culturing guidelines in abalone species and suggest the feasibility of culturing abalone ovary-dissociated cells.

Factors Affecting Primary Cultures of Haliotis discus hannai Ovary-dissociated Cells and General Culture Aspects

류준형,남윤권,공승표 한국수산과학회 2015 Fisheries and Aquatic Sciences Vol.18 No.1

We investigated factors affecting primary cultures of Pacific abalone Haliotis discus hannai ovary-dissociated cells to identify general aspects of their early-phase culture. Ninety-seven cell populations derived from 30 individuals were cultured in different media with varying compositions of medium supplements, and initial attachment, subculture, and survival for ≥10 weeks were assessed according to medium composition and individual. We also examined the time required for subculture and the rate of cell death according to both culturing period and passage number within 10 weeks. A lack of fetal bovine serum (FBS) and hemolymph significantly inhibited the growth of cultured cells, while we detected no significant effect of medium composition on initial cell attachment. Through data reallocation, with the omission of data from cell populations cultured in FBS-free and hemolymph-free media, we showed that growth inhibition was also affected by individual differences among the abalones used. During the culture, we observed four different types of cell morphology. Moreover, considerable time was required for subculture—18.4 and 19.5 days for first and second subcultures, respectively—and cell death did not occur within 30 days or for passage 0. Our results will provide valuable information for developing universal cell culturing guidelines in abalone species and suggest the feasibility of culturing abalone ovary-dissociated cells.

Original Article : Factors Affecting Primary Cultures of Abalone Haliotis discus hannai Ovary-dissociated Cells and General Culture Aspects

( Jun Hyung Ryu ),( Yoon Kwon Nam ),( Seung Pyo Gong ) 한국수산과학회(구 한국수산학회) 2015 Fisheries and Aquatic Sciences Vol.18 No.1

We investigated factors affecting primary cultures of Pacific abalone Haliotis discus hannai ovary-dissociated cells to identify general aspects of their early-phase culture. Ninety-seven cell populations derived from 30 individuals were cultured in different media with varying compositions of medium supplements, and initial attachment, subculture, and survival for ≥10 weeks were assessed according to medium composition and individual. We also examined the time required for subculture and the rate of cell death according to both culturing period and passage number within 10 weeks. A lack of fetal bovine serum (FBS) and hemolymph significantly inhibited the growth of cultured cells, while we detected no significant effect of medium composition on initial cell attachment. Through data reallocation, with the omission of data from cell populations cultured in FBS-free and hemolymph-free media, we showed that growth inhibition was also affected by individual differences among the abalones used. During the culture, we observed four different types of cell morphology. Moreover, considerable time was required for subculture.18.4 and 19.5 days for first and second subcultures, respectively.and cell death did not occur within 30 days or for passage 0. Our results will provide valuable information for developing universal cell culturing guidelines in abalone species and suggest the feasibility of culturing abalone ovary-dissociated cells.

누에 실샘 가수분해물의 제조 및 특성 규명

황정욱 ( Jung Wook Hwang ),이희삼 ( Heui Sam Lee ),김호진 ( Ho Jin Kim ),김규오 ( Kyu Oh Kim ),최용수 ( Yong Soo Choi ) 한국잠사학회 2012 한국잠사곤충학회지 Vol.50 No.2

Silk protein has been explored to be used for biomedical applications for several decades. However, it has not been used in this field cause to their irreversible crystallization after dissolving in water. The existing methods of silk protein hydrolysis using silkworm cocoon were used with harmful solvents and through a very complicated process. Therefore, we have developed novel methods for the production of water-soluble hydrolysate using silkworm gland. We manufactured two types of silkworm gland-derived hydrolysate (water-soluble SGH, SSGH; total SGH, TSGH) and compared the characteristics with commercial cocoon-derived sericin hydrolysate (CSH). The molecular weight of SGH ranged from 7 to 50 kDa (SSGH) and 5 to 15 kDa (TSGH) within glycine, alanine, and aspartic acid as a main amino acid composition. In contrast, CSH ranged from 15 to 50 kDa within serine and aspartic acid. The results of FTIR implied that SGH was more soluble form than CSH, as shown by the decrease in the β-sheet structure at 1630 cm-1 on amide I peak. In comparison with 10% fetal bovine serum, 0.1% (1 mg/ml) SSGH had equivalent effect on the proliferation of human dermal fibroblasts and mesenchymal stem cells. All results of the SSGH made by novel manufacturing process indicate the SSGH is more preferable as a culture medium supplement than cocoon-derived sericin.