Background: Although an understanding of the proliferation and differentiation of fish female germline stem cells (GSCs) is very important, an appropriate threedimensional (3D) research model to study them is not well established. As a part of the development of stable 3D culture system for fish female GSCs, we conducted this study to establish a 3D aggregate culture system of ovarian cells in marine medaka, Oryzias dancena. Methods: Ovarian cells were separated by Percoll density gradient centrifugation and two different cell populations were cultured in suspension to form ovarian cell aggregates to find suitable cell populations for its formation. Ovarian cell aggregates formed from different cell populations were evaluated by histology and gene expression analyses. To evaluate the media supplements, ovarian cell aggregate culture was performed under different media conditions, and the morphology, viability, size, gene expression, histology, and E2 secretion of ovarian cell aggregates were analyzed. Results: Ovarian cell aggregates were able to be formed well under specific culture conditions that used ultra-low attachment 96 well plate, complete mESM2, and the cell populations from top to 50% layers after separation of ovarian cells. Moreover, they were able to maintain minimal ovarian function such as germ cell maintenance and E2 synthesis for a short period. Conclusions: We established basic conditions for the culture of O. dancena ovarian cell aggregates. Additional efforts will be required to further optimize the culture conditions so that the ovarian cell aggregates can retain the improved ovarian functions for a longer period of time.

In response to the expanding landscape of the biotechnology industry and the increasing demand for comprehensive drug development as well as the conduct of preclinical and clinical trials, there is a growing need for employment of diverse animal models, including both small and large animals. The focus of this study was on refining ex vivo culture techniques for bioluminescence imaging following administration of intradermal injections in large animals. To examine the feasibility of our approach, varying concentrations of the rFluc protein were administered to rats and live imaging was employed to validate the corresponding levels of expression. Subsequently, following administration of rFluc to mini-pigs, ex vivo analyses were performed on sample tissues to assess the levels of protein expression across different concentrations. In particular, optimal culturing conditions that facilitated the sustained expression of the protein in samples post-euthanasia were identified. Moreover, by employing small animal imaging devices, we were able to capture clear images of the sample plates, which provided evidence of the successful application of our experimental techniques. The findings from this research represent a significant effort toward refining bioluminescence imaging methods tailored for use with large animal models—an imperative facet of contemporary drug development and biomedical research.

Protein is an essential nutrient for humans to sustain life, but it is predicted that it will be challenging to secure protein through the traditional livestock industry in the future. Microalgae has high future value as an alternative protein food source due to resource utilization and sustainability advantages. In order to increase productivity, the culture conditions of microalgae, Chlorella vulgaris, Dunaliella salina, and Scenedesmus obliquus were examined in this study. The optimal culture conditions of C. vulgaris were mixotrophic culture, 25oC culture temperature, 7.0 initial pH, 10% initial inoculation, stirring culture, 3000 Lux light intensity, and 24L:0D light/dark cycle period with red LED. For D. salina, the optimal culture conditions were mixotrophic culture, 20oC culture temperature, 8.0 initial pH, 10% initial inoculation, stirring culture, 6000 Lux light intensity, and 12L:12D light/dark cycle period with white LED. For S. obliquus, the optimal culture conditions were mixotrophic culture, 30oC culture temperature, 8.0 initial pH, 10% initial inoculation, stirring culture, 4500 Lux light intensity, and 14L:10D light/dark cycle period with fluorescent light. These findings can be used as important information for increasing the production of microalgae as an alternative protein material resource in the future.

프로바이오틱스 제품에 대한 수요가 지속적으로 증가하 고 있으며, Lactobacillus 균주가 가장 대중적인 프로바이 오틱스로 널리 사용되고 있다. 프로바이오틱스는 기준에 적합한 균수의 확보가 중요하며 제조원가나 시간 등을 낮 추기 위해 배양법의 개발이 필요하므로 Lactobacillus 생 산을 위한 배양 조건이 최적화되었다. 반응표면방법론에 의한 통계적 최적화에서 반응 변수에 영향을 미치는 독립 변수의 최적 조건은 Lactobacillus acidophilus의 경우 22.55 시간(배양시간), 25oC(배양온도), 3.41%(프리바이오틱스 농 도); Lactiplantibacillus plantarum의 경우 24시간, 30.86oC, 2.00%; Lacticaseibacillus rhamnosus의 경우 66.67시간, 35oC, 3.41%이었다. Lactobacillus의 최적 배양조건은 예측 한 결과와 실제 결과가 밀접하게 일치하는 것을 확인하였 다. 이러한 데이터는 수율 높은 Lactobacillus를 생산하는 데 중요한 포인트를 제공할 것이다.

한국에서 알스트로메리아(Alstroemeria)는 저온 작물로 난 방비 절감 효과가 있고 장식용 절화 소재로 꾸준히 소비되어, 일정 재배면적이 유지되고 있다. 본 연구는 알스트로메리아 ‘한에로스’의 기내 대량증식과 뿌리 발근 조건을 구명하고 무 균묘 생산 시스템을 구축하기 위해 수행했다. 기내 대량 증식 조건 구명 실험에는 Murashige and Skoog(MS) 배지(3% sucrose, 0.25% gelrite)에 BA 단용 처리(BA 0.25, 0.5, 0.75mg･L-1)와 NAA 0.2mg･L-1에 BA를 농도(BA 0.25, 0.5, 0.75mg･L-1)에 따라 혼용 처리하였다. 발근 조건 구명을 위 해 MS배지(3% sucrose, 0.25% gelrite)에 BA 0.25 mg･L-1 단용 처리와 BA 0.25mg･L-1에 NAA를 농도(NAA 0.1, 0.2, 0.3mg･L-1)에 따라 혼용 처리하였다. 뿌리줄기 증식 실험 결 과, 호르몬 첨가에 따른 생육차이는 뿌리줄기 수를 제외한 나 머지 부분에서 뚜렷하게 확인되었다. 단용 처리구 보다 혼용 처리구에서 많은 신초가 유도되었다. BA 0.5mg･L-1와 NAA 0.2mg･L-1 혼용 배지에서 가장 많은 1.43개의 신초가 유도되 었다. 발근 실험 결과, ‘한에로스’는 BA 0.25mg･L-1 와 NAA 0.3mg･L-1 혼용 배지에서 뿌리 수가 3.96개로 가장 많고, 뿌 리가 짧고 두꺼워 순화 과정 중에 뿌리 손실이 적었다. 또한, 순화 과정 중 묘가 고사하지 않았다. 종합적으로, ‘한에로스’ 는 BA 0.5mg･L-1와 NAA0.2 mg･L-1 혼용 배지에서 증식을 하는 것이 적절하며, BA 0.25mg･L-1와 NAA 0.3mg･L-1 혼 용 배지에서 발근 후 순화하는 것이 최적의 기내 배양 조건으 로 사료된다. 본 연구의 결과로 알스트로메리아 무균묘의 보 급 확대에 기여하고자 한다.

This study investigated the culture characteristics of Cryptoporus volvatus, whichis grow naturally in Korea, to determine the suitable environmental conditions for its cultivation. The physiological characteristics of the mycelia were assessed according to the cultivation conditions, to determine the optimal conditions for artificial cultivation. The visual characteristics of the hyphae of Cryptoporus volvatus KACC52303 included an irregular and uneven surface and a fuzzy or cotton-like texture. Under the microscope, its microstructure showed pre-chlamydospore formation, but no clamps were seen. The appropriate culture temperature was found to be a medium/high temperature of approximately 25–30oC, and the optimal pH was found to have a wide range from weakly acidic (pH 4) to neutral (pH 7). In the optimal nutrient source experiment, hyphal growth was shown to be fair in a mixed medium with 2.5% dextrin as the carbon source and 0.1% yeast extract as the organic nitrogen source. Among the various amino acids, organic acids, and inorganic salts tested, the fastest hyphal growth was observed in the presence of leucine, acetic acid or gluconic acid, and KCl or KH2PO4, respectively. The column test showed that the best mycelial growth occurred in a mixed medium of 80% pine sawdust, 10% rice bran, and 10% corncob sawdust.

Roridin E, produced by Podostroma cornu-damae, is a mycotoxin with anticancer activity. To increase the content of roridin E, submerged culture conditions were optimized using response surface methodology. Three factors, namely, medium initial pH, incubation time and agitation speed were optimized using a Box–Behnken design. The optimum submerged culture conditions to increase the content of roridin E included a medium with an initial pH of 4.0, an incubation time of 12.90 days, and an agitation speed of 63.03 rpm. The roridin E content in the submerged culture, under the aforementioned conditions, was 40.26 mg/L. The findings of this study can help lower the current price of roridin E and promote its related research.

The effects of supplementing ESP-FM (Erythrobactor sp.), freshwater Chlorella (Chlorella sp.), and baker’s yeast (Saccharomyces cerevisiae) on the nutritional value and mass production of Moina macrocopa, which is used as a live feed for fish fry production, was investigated. Consequently, the effects of feeding the enriched M. macrocopa to the nutritional composition of larval rockfish (Sebastes schlegel) and carp (Cyprinus carpio) was also investigated. Maximum density of M. macrocopa was reached within 15-21 days after inoculation (0.5 to 22 individual/mL), at various temperatures, and either decreased or remained almost constantly thereafter. Protein content and amino acids composition of M. macrocopa were found to be influenced by their respective diets while lipid and ash contents did not considerably change. M. macrocopa fed with baker’s yeast were low in n-3 HUFA content, and those fed on the freshwater chlorella were high in the 18:2n-6 and 18:3n-3 HUFA content, and in cultures treated with ESP-FM were high content in n-3 HUFA. The utilization of M. macrocopa as a substitute fish feed for carp and rockfish showed the enrichment nutritional content.

The purpose of this study is to optimize the composition of the medium for turmeric fermentation and to select competent turmeric fermentation strains using bacterial isolates from kimchi. Initially, 30 isolates from kimchi were cultured in 5% (w/v) yeast extract and 1% (w/v) maltodextrin to determine viability. As a result, eight strains showed a tendency to maintain viability until the fifth day of fermentation. Subsequently, the eight isolates were fermented in an optimum medium for turmeric fermentation, 5% (w/v) yeast extract, 1% (w/v) maltodextrin, and 5% (w/v) turmeric for seven days to determine the viable cell count and antioxidant capacity. The antioxidant capacities of turmeric fermented by the eight isolates were similar or higher than turmeric fermented by Lactococcus lactis KCTC 2013, while maintaining high viable cell counts of both the eight isolates and L. lactis KCTC 2013 until the seventh day of fermentation. The antioxidant capacities of the selected five strains during fermentation might increase possibly due to the biological conversion of active compounds in turmeric by fermentation. Consequently, a total of five strains of the isolates showing higher antioxidant capacity (4.81±0.19-5.81±0.04 VCE/mL) than fermentation day 0 were selected for fermentation of turmeric.

노란달걀버섯은(Amanita javanica) 국내에서 산림법으 로 보호받고 있는 식용 가능한 외생균근성 버섯이나, 중요한 산림자원으로서 활용하기 위한 기초적인 특성 연구에 대한 자료는 부족한 실정이다. 이에 본 연구는 국내 채집 야생버섯 자실체로부터 분리한 노란달걀버섯 NIFoS 1267 균주를 이용하여 PDA 배지 상에서 물리적 요인(온도, pH, 광)과 화학적인 요인(염분, 중금속, 농약)에 따른 균사생장 특성을 조사하였다. 최적의 물리적 환경은 온도가 30oC, pH가 5-6, 암조건으로 배양이었을 때 노란달걀 버섯 균주의 균사생장이 가장 좋은 것으로 나타났다. 화학적 요인으로서 염분은 2.0% 농도 조건까지 버섯균주의 균사 생장이 가능하였다. 50 ppm 농도의 중금속 이온 환경에서 비소(As) 이온은 노란달걀버섯 균주의 균사생장에 영향을 주지 않았으나 카드뮴(Cd)와 납(Pb) 이온은 균사 생장이 불가능하게 하였다. 국내 산림에 사용되고 있는 2가지 농약의 경우, Abamectin 첨가 배지에서는 노란달걀 버섯 균주의 균사 생장에 영향이 없었으나 Acetamiprid, Emamectin benzoate, Thiacloprid가 첨가된 배지 환경에서는 균사 생장이 모두 다 저해 되었다. 본 연구 결과는 향후 상업적 생산을 위한 새로운 자원으로서 노란달걀버섯의 인공재배 연구에 도움이 될 것으로 기대된다.

To improve the productivity of shiitake mushroom (Lentinula edodes), seven different types of media for liquid spawn (denoted as “A” to “G”) were prepared with 0.3% soybean meal and varying sugar and glucose concentrations. During 14 days of incubation, the pH of the liquid culture gradually acidified with increasing incubation period. Additionally, there was a significant, but not prominent, difference in the degree of acidification depending on the sugar to glucose ratio. Liquid spawn culture “G,” which had the highest sugar content was the most acidic on the last day of incubation. Mycelium dry weight increased significantly with increasing incubation period, and there was no significant difference in mycelium dry weight irrespective of the sugar to glucose ratio even after 14 days of culture. The inoculation of liquid spawn in sawdust medium with an inoculation volume ≥ 45 mL and incubation period of 15 to 18 days were the optimal culture conditions. Productivity of fruit bodies in sawdust medium and mushrooms treated with liquid spawn was significantly higher compared to solid spawn treatment. The mushrooms treated with liquid spawn had better chewiness, and the free amino acid content, which is associated with savory taste, was higher in these mushrooms compared to those treated with solid spawn.

느타리버섯의 생산성 향상을 위하여 액체종균 배양액의 조건을 설정하고자 하였다. Potato dextrose 배양액에서 균체 생장을 위한 최적 pH는 6이었다. 대두박 0.3%와 설탕 및 포도당 함량을 달리한 15가지의 배양액(‘A’~‘O’)은 22℃및 pH 6의 조건에서 14일동안 배양되었을 때 설탕과 포도당이 각각 함유된 배양액에서 흡광도는 증가되었다. 특히 5~20%의 포도당만 함유된 배양액(‘C’, ‘D’, E’ 및 ‘F’)은 배양기간의 경과에 따라 흡광도가 증가되는 경향이었다. 배양액의 건조 균체량은 20%이하의 설탕이나 포도당이 단독으로 함유된 배양액에서 감소되었으나, 30%의 설탕(‘A’) 또는 포도당(‘B’)이 각각 함유된 배양액에서 건조 균체량은 증가되었다. 1100 mL의 톱밥배지 병재배 시 액체종균의 최적 접종량은 15~20 mL이었다. 고체종균 및 액체종균으로 느타리버섯을 재배하여 조직감을 비교한 결과 고체종균에 비해 액체종균으로 재배된 버섯에서 우수한 경향이었다.

In order to develop solid mass cultivation technology that maximizes insecticide of entomopathogenic nematode(EPN) isolated from Korea, we have studied the optimization of solid culture medium and production conditions. The optimized conditions from the harvest yields and the insecticidal activity against the Galleria mellonella larvae were yeast extract 2.0%, soybean flour 20%, whole milk powder 3%, olive oil 5% and egg yolk 5%. It was also found that the optimum condition of the carrier(polyurethane) content was about 8-12%, the culture time of symbiotic bacteria was about 48 hours, and the inoculation concentration of nematode was 4,000-5,000 per g medium. And the optimal harvesting point in solid culture conditions was confirmed after 12 days of inoculation.

In the last several decades, cell therapy research has increased worldwide. Many studies have been conducted on cell therapy, and have revealed that transplanted cells did not survive for long, and implanted cells remained inactive causing immune rejection depending on the patient’s condition. Therefore, studies on cell-free therapy need to be conducted. To overcome these limitations, an alternative is the use of supernatant from cells, called “conditioned media (CM).” During in vitro cell culture, culture media supply nutrients to maintain cell characteristics and viability. In the culture, cells not only consume nutrients but also release beneficial proteins and substances, which are called “secretome.” CM from cells can be stored for a long time and is easy to handle. Moreover, secretome in CM can also be measured; exact amount of secretome is important to set the standard value for disease treatment. Here, we reviewed studies on CM and confirmed that various secretomes from CM were identified in these studies. Moreover, these findings could benefit cell and animal studies in future. In conclusion, CM could be a potential candidate for an alternative to cell therapy.

Streptomyces species have been studied to find potent pest control agents as an alternatives to chemical insecticides.Previously, one of the ethyl acetate extract of Streptomyces isolates cultured on unpolished rice medium showed highlevel of juvenile hormone antagonist and larvicidal activities against pest insects including Aedes albopictus, Plutella xylostellaand Laodelphax striatellus. It has been known that the biosynthesis of secondary metabolites of Streptomyces could beinfluenced by a variety environments such as nutritional composition and growth conditions. In this study, to optimizeculture conditions for stable production of insecticidal compounds from this isolate, binding assay and bioassay-guidedmonitoring were conducted using various culture conditions.

반하(Pinellia ternata(Thunb.) Breit)는 천남성과에 속하는 다년생 초본식물로서 조직배양을 이용하 여 대량번식 방법 연구가 활발히 진행되어 왔다. 하지만 대량생산된 괴경들의 토양 순화 및 적응을 위 한 환경 조건의 확립의 연구는 미비하다. 따라서 본 연구에서는 기내에서 증식된 반하의 괴경을 이용하 여 우수한 품질의 약재와 건전묘의 대량생산을 위해 생육에 적합한 토양조건을 탐색하였다. 본 연구에 서는 액체배지에서 현탁배양 한 괴경(Type 1)과 고체배지에 치상하여 배양된 괴경(Type 2)두 종류를 비 교하였다. 토양은 3개의 조성으로 조합하여 생육을 비교하였으며, 코코피트, 피트모스, 버미큘라이트, 펄라이트 및 제오라이트를 배합하여 사용하였다. 기내 배양 조건이 다른 처리구들의 토양 조성별 순화 율 측정하였으며, 생육의 차이를 확인하기 위해 8주동안 생육 후 초장, 잎 수, 마른잎 수, 괴경 수, 괴 경 크기, 생체중 및 건물중을 측정하였다. 또한 주아의 생성율을 확인하기 위하여 4주와 8주에 측정을 진행하였다. 그 결과 Type 2가 펄라이트를 20%증량한 상토 B에서 가장 우수한 생육을 보였다. 괴경의 비대에 영향을 주고 묘로서 이용을 위하여 필요한 잎의 수는 상토 B에서 가장 많은 1.7개로 나타났고 가장 잎의 출현이 없었던 Type 1의 상토 C가 0.9개로 나타나 1.8배의 차이를 나타냈다. 또한 같은 처 리구에서 건물중이 통계적으로 유의한 차이를 보이며 우수한 것으로 나타났다. 반하의 주아 생성율의 차이는 상토 B에서 1.1개로 우수하였으며, 가장 저조한 처리구의 수치보다 1.2배 높은 생성율을 확인할 수 있었다. 이러한 결과로 기내에서 배양된 반하의 토양 순화 및 적응기에 괴경의 비대를 유도하여 우 수한 한약재 생산이 가능 할 것으로 생각되며 차후 토양에서 재배 시 반하의 대량번식에도 도움이 될 것이라 사료된다.