• Title/Summary/Keyword: culturing

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Periodontal healing using a collagen matrix with periodontal ligament progenitor cells in a dehiscence defect model in beagle dogs

  • Yoo, Seung-Yoon;Lee, Jung-Seok;Cha, Jae-Kook;Kim, Seul-Ki;Kim, Chang-Sung
    • Journal of Periodontal and Implant Science
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    • v.49 no.4
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    • pp.215-227
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    • 2019
  • Purpose: To histologically characterize periodontal healing at 8 weeks in surgically created dehiscence defects in beagle dogs that received a collagen matrix with periodontal ligament (PDL) progenitor cells. Methods: The bilateral maxillary premolars and first molars in 6 animals were used. Standardized experimental dehiscence defects were made on the buccal side of 3 premolars, and primary culturing of PDL progenitor cells was performed on the molars. Collagen matrix was used as a scaffold and a delivery system for PDL progenitor cells. The experimental sites were grafted with collagen matrix (COL), PDL progenitor cells with collagen matrix (COL/CELL), or left without any material (CTL). Histologic and histomorphometric analyses were performed after 8 weeks. Results: The defect height from the cementoenamel junction to the most apical point of cementum removal did not significantly differ across the CTL, COL, and COL/CELL groups, at $4.57{\pm}0.28$, $4.56{\pm}0.41$, and $4.64{\pm}0.27mm$ (mean ${\pm}$ standard deviation), respectively; the corresponding values for epithelial adhesion were $1.41{\pm}0.51$, $0.85{\pm}0.29$, and $0.30{\pm}0.41mm$ (P<0.05), the heights of new bone regeneration were $1.32{\pm}0.44$, $1.65{\pm}0.52$, and $1.93{\pm}0.61mm$ (P<0.05), and the cementum regeneration values were $1.15{\pm}0.42$, $1.81{\pm}0.46$, and $2.57{\pm}0.56mm$ (P<0.05). There was significantly more new bone formation in the COL/CELL group than in the CTL group, and new cementum length was also significantly higher in the COL/CELL group. However, there were no significant differences in the width of new cementum among the groups. Conclusions: PDL progenitor cells carried by a synthetic collagen matrix may enhance periodontal regeneration, including cementum and new bone formation.

Establishment of efficient Alstromeria callus induction system using node culture and various hormones (마디배양과 다양한 호르몬을 이용한 효율적인 알스트로메리아 캘러스 유도 시스템 체계 확립)

  • Yang, Hwan Rae;Lee, Sang Hee;Kim, Jong Bo
    • The Journal of the Convergence on Culture Technology
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    • v.5 no.1
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    • pp.413-416
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    • 2019
  • Alstroemeria (Alstroemeriaceae) is one of the most important cut flowers in international market. Especially, characteristics like long vase-life, various colors, tolerance to low temperature and a low energy requirement during cultivation have stimulated this success. Because of its characteristics such as low multiplication rates, time-consuming process and high risk of carrying viral disease, in vitro propagation techniques based on rhizome meristems culture have been developing nowadays. The callus induction has various cultivation sites compared with the direct plant generation method, and if the callus is maintained well, the plant differentiation can be performed simultaneously while maintaining the callus, so that it can be used for mass proliferation. In this study, we tested various hormones and cultivars for efficient callus induction. As a result of culturing between the nodes and the internodes, the callus began to be formed after 8 weeks, and the calli incidence in the nodes was higher than that between the internodes. Also, in the comparison of 2,4-D and picloram, the callus incidence rate was up to 2 times higher in the medium treated with 2,4-D. Using these results, it is thought that it will help establish the system of mass propagation system of Alstroemeria and cultivate new varieties.

Dynamics of fungal community during silage fermentation of elephant grass (Pennisetum purpureum) produced in northern Vietnam

  • Vu, Viet Ha;Li, Xiyang;Wang, Mengyuan;Liu, Rongmei;Zhang, Guojian;Liu, Wei;Xia, Baixue;Sun, Qun
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.7
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    • pp.996-1006
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    • 2019
  • Objective: This study aimed to gain deeper insights into the dynamic changes in spoilage fungi populations during fermentation and the influence of traditional additives on silage quality. Methods: Elephant grass (Pennisetum purpureum) was prepared without any additive (control), and with the addition of 0.5% salt, and 0.5% salt-0.2% sugar mixture. The fungal community was then determined using a classic culturing method and high-throughput sequencing at 0, 5, 15, and 60 days after ensiling. Results: The results showed that the fungal community of elephant grass silage varied significantly between the natural fermentation without any additive and the two additive groups. The diversity and relative abundance of spoilage molds in the control group were much higher than those in the two treatment groups (p<0.05). Three species of yeasts (Candida sp., Pichia sp., Trichosporon sp.) and four spoilage molds (Fusarium sp., Aspergillus sp., Muco sp. and Penicillin sp.) were the predominant fungi in elephant grass during natural fermentation from 0 to 60 days, which were found to be significantly decreased in salt and sugar additive groups (p<0.05). Meanwhile, the diversity and relative abundance of undesirable molds in the 0.5%-salt additive group were the lowest among all groups. Conclusion: Adding salt and sugar, particularly 0.5% salt, is a promising effective approach to reduce the amount of undesirable fungi thus, improving the silage quality of elephant grass in northern Vietnam.

Effect of sodium chloride on the growth, amino acid content, and fragrance patterns of Pleurotus ostreatus (염화나트륨 농도가 느타리 자실체의 아미노산과 향기성분에 미치는 영향)

  • Lee, Sang-Chul;Pyeon, Ha-Young;Park, Youn-Jin;Oh, Tae-Seok;Jang, Myoung-Jun
    • Journal of Mushroom
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    • v.19 no.4
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    • pp.310-315
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    • 2021
  • We investigated the effect of sodium chloride-associated abiotic stress on the development of Pleurotus ostreatus. We examined the growth characteristics of fruiting bodies, constituent amino acids, and fragrance pattern to determine the effect of culturing Pleurotus ostreatus on a sawdust substrate supplemented with sodium chloride in a dose-dependent manner. Pleurotus ostreatus fruiting bodies exhibited an increasing tendency towards augmented yields when grown in the presence of 0.5% sodium chloride as compared with that grown in the control group. However, increasing the supplementation of sodium chloride from 1.0 % to 2.0% resulted in significantly decreased yields of Pleurotus ostreatus fruiting bodies in these groups as compared with that in control groups. Further assessment revealed the presence of 14 types of amino acids in the fruiting bodies, including aspartate, threonine, serine, glycine, alanine, methionine, valine, isoleucine, leucine, phenylalanine, tyrosine, lysine, histidine, and arginine, at lower levels in all the sodium chloride-treated groups than in the control group; except for glutamic acid and proline. Similarly, fragrance pattern analysis of the Pleurotus ostreatus fruiting body by chromatography confirmed that the intensity of the substances presumed to be octane compounds, to which the unique flavor of mushrooms is attributed, was lower in all the sodium chloride-treated groups than in the control group.

Transcriptomic Analysis of the Difference of Bovine Satellite Cell Between Longissimus dorsi and Semimembranosus on Hanwoo Muscle Tissues (한우의 등심과 사태조직 유래 근육위성세포의 성장단계별 유전발현 차이 분석)

  • Kim, H.J.;Kang, D.H.;Park, B.H.;Lee, W.Y.;Choi, J.H.;Chung, K.Y.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.23 no.1
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    • pp.117-128
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    • 2021
  • The skeletal muscle development of Hanwoo steer has been processed in the prenatal and postnatal periods. Bovine satellite cell located in perimysium of muscle tissues has differentially distributed in peripheral tissues. The study of postnatal development of satellite cells can help understand the genetic and functional regulation of meat characteristics. Factors affecting muscle size increase are related to the accumulation of DNA or synthesis of RNA proteins. In this study, we observed muscle development and differentiation after culturing bovine satellite cells derived from longissimus dorsi and semimembranosus regions of Hanwoo muscle tissue. In addition, RNA sequencing data were analyzed for differentially expressed genes (DEG) involved in intracellular muscle development and growth. The DEG of the two muscle tissues were compared according to 1day, 2day, 4day, and 7day. The overall gene expression level was confirmed by the heat map. Gene Ontology (GO) classification method was used to compare the expression level of gene groups affecting LD and SM development. The histology of GO was consistent with the time-cause change of LD and SM cell morphology. SM showed more active skeletal muscle development than LD. Even within the same time, SM expressed more genes than LD, thus synthesizing more muscle fibers

An Analysis of the Current Status and Characteristics of Airborne Fungi in Indoor Air in Multi-Use Facilities Nationwide (전국 다중이용시설의 실내공기 부유 곰팡이의 현황 및 특성 분석)

  • Park, Yongsung;Kwon, Soonhyun;Park, Song-Yi;Kee, Sun-Ho;Yoon, Wonsuck
    • Journal of Environmental Health Sciences
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    • v.48 no.5
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    • pp.282-289
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    • 2022
  • Background: Airborne fungi are ubiquitous in the air and exposure to an airborne fungus can be a significant risk factor. The composition of fungi has been potentially important for human health, especially for respiratory diseases like asthma and atopic dermatitis. Therefore, we attempted to ascertain what kind of airborne fungi affect human health at a nationwide level. Objectives: This study was carried out to provide information on indoor fungi distribution at multi-use facilities throughout South Korea. Methods: We classified our data by region and public facility after collection, cultivation, and identification via the sequencing of the ITS (internal transcribed spacer) region. We investigated whether or not the proliferation of HaCaT cells was affected by the identified airborne fungi. Results: In our data, the most isolated airborne fungi by region were Penicillium spp (Seoul, Daegu), Periconia sp (Gyeonggi-do), Iprex sp (Gangwon-do), Phanerochaete sp (Busan), Bjerkandera sp (Gwangju), and Aspergillus sp (Jeju-do). In the public facilities, the most detected fungi were Cladosporium sp (public transport), Penicillium sp (apartment house, retail market, financial institution, karaoke room), Bjerokandera sp (underground parking lot, public toilet, medical institution), Periconia sp (retail store), and Fusarium sp (general restaurant). Next, we selected twenty airborne fungi to examine their cytotoxicity and proliferation of human skin cells. In this experiment, the proliferation of the cells was influenced by most of the identified fungi. In case of the cytotoxicity test, most genera except for Rhodotorula sp and Moesziomyces sp showed cytotoxicity in HaCaT cells. Conclusions: The distribution of mold in the indoor air in multi-use facilities in South Korea differs from region to region, and this is an indicator that should be considered in future health impact studies. In addition, as a result of culturing about 20 types of bacteria dominant in indoor air, it was found that most (90%) inhibit the growth of skin cells, which can be harmful to health. An in-depth study of the health effects of floating fungi is needed.

A simple mid-term preservation method (SMPM) of plant callus under low temperature conditions (저온 보존을 이용한 간편 중기 식물캘러스 저장법)

  • Park, Sung-Chul;Park, Su Hyun;Kim, Soyoung;Jeong, Yu Jeong;Kim, Cha Young;Jeong, Jae Cheol
    • Journal of Plant Biotechnology
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    • v.49 no.3
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    • pp.187-192
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    • 2022
  • The repeated monthly or weekly subculture of plant callus is labor intensive and increases the risk of somaclonal variation from the parental callus line. The most effective method for preserving plant callus is cryopreservation, which involves storage in liquid nitrogen. However, this method cannot be applied to the callus of different plant species in the same manner, so it is difficult to develop a standardized cryopreservation method. In addition, the survival rate of the frozen callus after thawing and the regeneration rate after survival are uncertain. Therefore, it is necessary to develop a method to extend the subculture interval of plant callus in an active state. In this study, active plant calli of various species without freezing was incubated at 15℃ for 4 to 12 weeks without subculture. After 12 weeks, 8 lines of plant callus grew less than 2-fold when cultured at 25℃, but at least 2 times as much when cultured at 15℃. Moreover, total antioxidant activity did not differ significantly between plant callus recovered at 25℃ after culturing at 15℃ or at 25℃. These results show that the subculture interval can be extended at a temperature of 15℃ without need for modified medium composition or additional processes. In addition, positive results in all calli of several plant species are expected to reduce labor as well as somaclonal variation by increasing the subculture.

Effects of different culture systems on the culture of prepuberal buffalo (Bubalus bubalis) spermatogonial stem cell-like cells in vitro

  • Li, Ting-Ting;Geng, Shuang-Shuang;Xu, Hui-Yan;Luo, Ao-Lin;Zhao, Peng-Wei;Yang, Huan;Liang, Xing-Wei;Lu, Yang-Qing;Yang, Xiao-Gan;Lu, Ke-Huan
    • Journal of Veterinary Science
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    • v.21 no.1
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    • pp.13.1-13.14
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    • 2020
  • Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3+ subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.

Breeding a new variety of white Hypsizygus marmoreus 'Baekmaru' with excellent cultivation stability and storability (재배적 안정성과 저장성이 우수한 백색 느티만가닥버섯 신품종 '백마루' 육성)

  • Oh, Youn-Lee;Kim, Minseek;Jang, Kab-Yeul;Oh, Min Ji;Im, Ji-Hoon;Lee, Jong-Won
    • Journal of Mushroom
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    • v.20 no.3
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    • pp.153-162
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    • 2022
  • The purpose of this study was to breed a variety of stable productivity and high storage characteristics of white Hypsizygus marmoreus, which has high demand at domestic and global market due to a low bitter taste and the preference for white species. Accordingly, 'Baekmaru' was bred by hybridizing brown and white cap color species and backcrossing with white species. The 'Baekmaru' variety was bred by crossbreeding the brown and white species and backcrossing the white species. Through repeated cultivation of 'Baekmaru', a variety with a low contamination rate was selected when culturing the spawn for stable cultivation. As a results of demonstration test, the yield of 'Baekmaru' was 14% higher than that of the commercial variety. In addition, among the fruiting body characteristics of 'Baekmaru', diameter and thickness of the pileus were 16.43±15.27mm and 6.46±0.58mm, which were slightly higher than the commercial variety, and the hardness was 2.69±0.89N for the pileus and 3.09±0.89N for the stipe. The shelf life showed less change in thickness and color of pileus compared to commercial variety. The hardness of pileus of 'Baekmaru' was maintained in the range of 3.5 to 4.0N in the 4℃ and 4℃ and 20℃ mixed treatment until 30 days of storage, and was higher than that of the commercial variety. Therefore, it was suggested to be excellent in storability.

Cultivation characteristics and yield of Sparassis latifolia depend on the substrate mixture (꽃송이버섯(Sparassis latifolia) 생육배지 조성에 따른 재배특성 및 수량)

  • Heo, Byong-Soo;Choi, Kyu-Hwan;Jo, Yeong-Min
    • Journal of Mushroom
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    • v.20 no.2
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    • pp.61-68
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    • 2022
  • We investigated the standard cultivation substrate for Sparassis latifolia "Neowul" bred in Jeollabuk-do Agricultural Research and Extension Services. Cultivation characteristics and yield were assessed by varying the kind of sawdust and additives, and the mixing ratio. The cultivation period in larch sawdust was the shortest at 97 days. The yield was excellent (143.6 g). The findings indicated that larch is a tree species appropriate for the cultivation of S. latifolia. When the additives were varied, the yield and productivity (53.1%) were highest (116.6 g) for the wheat bran treatment. Thus, wheat bran would be an additive appropriate for culturing S. latifolia. Concerning the use of different mixing ratios, larch sawdust:beet-pulp:wheat bran ratios of 80:15:5 and 85:10:5 resulted in yields of 114.4 g and 111.4 g, and productivity of 52.5% and 51.8%, respectively. These yield and productivity values were not statistically different. Thus, the standard cultivation substrate for S. latifolia can comprise larch sawdust, beet pulp, and wheat bran at a ratio of 80:15:5 or 85:10:5. The carbon/nitrogen (C/N) ratio assumed to be appropriate for the cultivation of S. latifolia was 184-223. Pinheading would be difficult below a C/N substrate ratio of 105. Thus, the C/N ratio of the media, as well as the pH, would be vital factors affecting pinheading during S. latifolia cultivation.