• Title/Summary/Keyword: In Vitro Development

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Effects of Recipient Oocytes and Electric Stimulation Condition on In Vitro Development of Cloned Embryos after Interspecies Nuclear Transfer with Caprine Somatic Cell (수핵난자와 전기적 융합조건이 산양의 이종간 복제수정란의 체외발달에 미치는 영향)

  • 이명열;박희성
    • Reproductive and Developmental Biology
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    • v.28 no.1
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    • pp.21-27
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    • 2004
  • This study was conducted to investigate the developmental ability of caprine embryos after somatic cell interspecies nuclear transfer. Recipient bovine and porcine oocytes were obtained from slaughterhouse and were matured in vitro according to established protocols. Donor cells were obtained from an ear-skin biopsy of a caprine, digested with 0.25% trypsin-EDTA in PBS and primary fibroblast cultures were established in TCM-199 with 10% FBS. The matured oocytes were dipped in D-PBS plus 10% FBS + 7.5 $\mu$ g/ml cytochalasin B and 0.05M sucrose. Enucleation were accomplished by aspirating the first polar body and partial cytoplasm which containing metaphase II chromosomes using a micropipette with an out diameter of 20∼30 $\mu$m. A Single donor cell was individually transferred into the perivitelline space of each enucleated oocyte. The reconstructed oocytes were electric fusion with 0.3M mannitol fusion medium. After the electrofusion, embryos were activated by electric stimulation. Interspecies nuclear transfer embryos with bovine cytoplasts were cultured in TCM-199 medium supplemented with 10% FBS including bovine oviduct epithelial cells for 7∼9 day. And porcine cytoplasts were cultured in NCSU-23 medium supplemented with 10% FBS for 6 ∼8 day at $39^{\circ}C, 5% CO_2 $in air. Interspecies nuclear transfer by recipient bovine oocytes were fused with electric length 1.95 kv/cm and 2.10 kv/cm. There was no significant difference between two electric length in fusion rate(47.7 and 44.6%) and in cleavage rate(41.9 and 54.5%). Using electric length 1.95 kv/cm and 2.10 kv/cm in caprine-porcine NT oocytes, there was also no significant difference between two treatments in fusion rate(51.3 and 46.1%) and in cleavage rate(75.0 and 84.9%). The caprine-bovine NT oocytes fusion rate was lower(P<0.05) in 1 pulse for 60 $\mu$sec(19.3%), than those from 1 pulse for 30 $\mu$sec(50.8%) and 2 pulse for 30 $\mu$sec(31.0%). The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(53.3%) and 2 pulse for 30 $\mu$sec(50.0%), than in 1 pulse for 60 $\mu$sec(18.2%). The caprine-porcine NT oocytes fusion rate was 48.1% in 1 pulse for 30 $\mu$sec, 45.2% in 2 pulse for 30 $\mu$sec and 48.6% in 1 pulse for 60 $\mu$sec. The cleavage rate was higher(P<0.05) in 1 pulse for 30 $\mu$sec(78.4%) and 1 pulse for 60 $\mu$sec(79.4%), than in 2 pulse for 30 $\mu$sec(53.6%). In caprine-bovine NT embryos, the developmental rate of morula and blastocyst stage embryos were 22.6% in interspecies nuclear transfer and 30.6% in parthenotes, which was no significant differed. The developmental rate of morula and blastocyst stage embryos with caprine-porcine NT embryos were lower(P<0.05) in interspecies nuclear transfer(5.1%) than parthenotes(37.4%).

Development of Vivorium, a new indoor horticultural ornamental plants via plant tissue culture techniques (식물조직배양 기술을 이용한 새로운 실내 원예 장식품인 비보리움(Vivorium)의 개발)

  • Hwang, Min Hee;Kim, Do Yeon;Cho, In Sun;Kim, Mi Hyung;Kwon, Hyun Sook;Kim, Jong Bo;Kim, Su Jung;Kim, Sun Hyung
    • Journal of Plant Biotechnology
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    • v.48 no.3
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    • pp.179-185
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    • 2021
  • Indoor gardening includes wall greening, terrariums, and flower arrangements. Among these types of indoor gardens, the terrarium is easy to access for the general public, but in Korea, because of the focus on esthetics, the original purpose of creating terrariums, which was to grow plants sustainably in an enclosed space, has been lost. In addition, miniaturization of plants is required to grow plants in an enclosed space. Since the available plant species suitable for a terrarium are limited, only plants such as succulents, cacti, and moss have been used. In this study, Bronze (X Graptosedum) was used, and these problems were solved using the following three methods: placement and growth of virus-free plants in the terrarium; extending the diversity of plants with minimal size that can be planted in terrariums; and reducing the price of in vitro plants with minimal size by achieving large-scale production. In particular, tissue-cultured succulents were developed into a Vivorium by replacing the tissue culture container and renewing the composition of the plant. This paper suggests a new indoor horticultural field, Vivorium, that can improve the current limitations of terrariums and make them more accessible to the general public. The introduction and popularization of new indoor gardening fields with the increase in single-person households and indoor activities in the Pandemic era can also improve psychological stability among people and in the society.

Study on skin anti-inflammatory activity of fig (Ficus carica L.) fruit extract fractions (무화과(Ficus carica L.) 열매 추출 분획의 피부 항염증 활성 연구)

  • Hee Joon Kwon;Geun soo Lee;Jin Hwa Kim;Soon Woo Kwon;Hyung seo Hwang
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.416-423
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    • 2023
  • Figs has known to have antioxidant, whitening, anti-inflammatory, and antibacterial effects in their leaves, roots, stems, latex, and fruits. In order to develop cosmetic materials based on natural products, we have studied on the skin activity of the ficin in latex as well as the whitening function of the fruit extract with 70% ethanol, and used it as a raw material for released cosmetic product. However, there is little research on the demand for the development of new eutectic solvent extraction methods and its ability to control skin inflammation and psoriasis regulation. Thus, in this study, we evaluated the effectiveness of fig fruit extracts and fractions using eutectic solvent extraction for skin inflammation control and psoriasis. First, fig fruits were extracted under optimal eutectic solvent conditions and fractionated with n-hexane, dichloromethane, ethyl acetate, and butanol. First, the antioxidant activity and inhibition of nitric oxide (NO) production were confirmed in mouse macrophage RAW264.7 cells. In addition, as a result of observing the mRNA expression through RT-PCR, pro-inflammatory cytokines such as TNF-α, IL1α, and IL-1β were suppressed significantly in the hexane, dichloromethane, and ethyl acetate fractions. In addition, it was confirmed in TNF-α stimulated HaCaT keratinocyte model. Finally, chemokine CC motif ligand 20 (CCL20), marker gene of human psoriasis skin disease, was significantly suppressed in the hexane, dichloromethane, and ethyl acetate fractions. These results suggested its anti-inflammatory and skin soothing effect and the possibility of development as an excellent skin soothing natural cosmetic material in the future through future clinical trials.

Effects of Anabolic Steroids of Pork on Proliferation and Differentiation of Myogenic Satellite Cell (돼지 고기의 아나볼릭 스테로이드가 Myogenic Satellite Cell의 증식과 분화에 미치는 영향)

  • Lee, Dong-Mok;Lee, Ki-Ho;Cheon, Yong-Pil;Chun, Tae-Hoon;Choi, In-Ho
    • Food Science of Animal Resources
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    • v.30 no.5
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    • pp.842-850
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    • 2010
  • Sex steroids are known to be involved in skeletal muscle development (anabolic effect) and are frequently used in medicines. It has been known that pork contains a variety of steroids that are mainly synthesized in the gonads (testis and ovary). Thus, the present study was conducted to evaluate the effects of anabolic steroids of pork on the proliferation and differentiation of myogenic satellite cells (MSC). Three different methods (M1, M2, and M3) were developed for the isolation and purification of steroids from porcine tissues. Among three extraction methods that we developed, M3 was the best method with respect to the quantities of steroids and the induction of MSC proliferation. Hormonal analysis showed that the steroid hormone levels were the highest in muscle and fat of intact male than those of castrated males and females. In addition, the highest serum levels of nandrolone and testosterone were detected in intact males, whereas estrone and $17{\beta}$-estradiol levels were similar in the entire experimental serum samples. Expression of androgen receptor (AR), myoD, desmin, and myogenin in bovine muscle cells were significantly up-regulated by the treatment of steroid extracts. The highest increas of myogenin and AR mRNA abundance were observed in the MSCs treated with M3 extract (p<0.001). Altogether, the present research showed the positive effect of steroids on MSC proliferation and differentiation in vitro. These results would certainly imply a beneficial effect of pork consumption on human muscle development.

Enhancement of the germination efficiency of Gastrodia elata seeds using a new Mycena species (신규 애주름버섯 NIFOS101을 이용한 천마 종자의 발아효율 증진)

  • Lee, Hyo-Ryeon;Han, Muho;Choi, Mi Na;Lee, Hyoshin;Lee, Seok-Woo;Park, Eung-Jun
    • Journal of Plant Biotechnology
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    • v.44 no.1
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    • pp.56-60
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    • 2017
  • This study investigated the in vitro symbiotic seed germination of the achlorophyllous orchid, Gastrodia elata, using a new Mycena species. A leaf-disc ($2{\times}2cm$) of Quercus accutissima was inoculated with either of the two fungal species, NIFOS101 (NCBI accession number KY449288) or KFRI1212 (HQ662845), following which it was placed on water agar medium, prior to sowing seeds of G. elata. After 20 days of symbiotic culture, NIFOS101 and KFRI1212 germinated 94% and 70% of G. elata seeds, respectively, although the initiation of protocorm development was induced much earlier by KFRI 1212 than by NIFOS101. Furthermore, the NIFOS101 mycelia grew much faster than KFRI 1212 at all temperatures tested. A phylogenetic analysis using the internal transcribed spacer (ITS) sequences showed that NIFOS101 belonged to a clade with M. purpureofusca, which completely differed from the clade with KFRI1212. This study not only identified a new fungal species, NIFOS101, which improved the rate of symbiotic seed germination up to 94% as compared to KFRI1212 (70%), but also revealed that G. elata required a broad taxonomic range of fungi for its symbiotic germination.

Quality and Antioxidant Activity of Yanggaeng Containing Herbal Medicine Extracts for the Elderly (약재 추출물을 첨가한 노인용 기능성 양갱의 품질 특성 및 항산화 효과)

  • Jeong, Se-Hyun;Kim, Jung-Hoan;Yang, Soo Jin;Lee, Sung-Hyen;Oh, Jung-Hoon;Lee, Jung Ok;Lee, Hae-Jeung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.9
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    • pp.1304-1310
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    • 2015
  • The purpose of this study was to investigate antioxidant activity of yanggaeng containing herbal medicine extracts (YH) by employing various in vitro antioxidant assays for 1,1-diphenyl-2-picrylhydrazyl free radical scavenging, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) free radical scavenging, and reducing power. We also investigated moisture content, pH, texture profile, and sugar content, and evaluated the sensory characteristics of developed yanggaengs. Moisture content of YH was 41.54%. Sugar contents increased while pH decreased as content of herbal medicine extracts increased. Lightness was not significantly different between YH and control. However, herbal medicine extracts enhanced the redness of yanggaeng. Texture profile analysis showed that gumminess and chewiness of yanggaeng added with herbal medicine extracts decreased compared to control. Herbal medicine extracts enhanced yanggaeng's antioxidant activity, reducing power, and scavenging ability. Herbal medicine extracts enhanced antioxidant properties of yanggaeng and made it easier to swallow.

Hypocholesterolemic Effect of Lyophilized, Heat-Killed Lactobacillus rhamnosus and Lactobacillus plantarum (가열살균한 Lactobacillus rhamnosus와 Lactobacillus plantarum의 콜레스테롤 저하 효과)

  • Kim, Dae-Weon;Yang, Dae-Hyeok;Kim, Sun-Young;Kim, Kwang-Soo;Chung, Myun-Gjun;Kang, Sang-Mo
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.69-74
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    • 2009
  • Lactic acid bacteria (LAB) were well known to enhance the intestinal health of human. For the development of pharmaceutical LAB. it was screened that the LAB with activity lowering the cholesterol in vitro and evaluated the hypocholestrolemic effect of live and heat-killed (HK) LAB on rats. The selected Lactobacillus plantarum CBT 1209 and Lactobacillus rhamnosus CBT 1702 had the deconjugation of bile salts and assimilation of cholesterol micelles activities from laboratory media, The mixture of 1702 and 1209 strains was administrated to the rats with high cholesterol diet. The experiment performed by 4 groups which were control, HCD, LLAB, HKLAB groups. The hypocholesterolemic effect of LAB (strains 1702, 1209) at blood level, the phenomena of AI decreasing through LDL-cholesterol dwindling, was assessed. This effect of 1702 and 1209 was enhanced when it comes to be the HKLAB more the live-LAB, This data means that the Lactobacillus rhamnosus CBT 1702 and Lactobacillus plantarum CBT 1209 were very useful functional ingredient for hypercholesterolemia.

A Study on the Development of Cosmetic Materials Using Unripe Peaches Seed Extracts (복숭아 유과 씨 추출물을 이용한 화장품 소재 개발에 관한 연구)

  • Kim, Da-Mi;Kim, Kyoung-Hee;Kim, Young-Shik;Koh, Jong-Ho;Lee, Kyong-Haeng;Yook, Hong-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.1
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    • pp.110-115
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    • 2012
  • Various unripe peach (Prunus persica L. Batsch) seeds, which were picked on late May, were subjected to 80% methanol extracts to investigate the antioxidant activities, whitening effects, and anti-wrinkle activities. The yield of 6 cultivars (Takinosawa Gold, Kawanakawase Hakuto, Madoka, Yumefuji, Nagasawa Hakuho, and Hong Bak) from unripe peaches were the highest on Yumefuji, followed by Madoka, Kawanakawase Hakuto, Takinosawa Gold, Hong Bak, and Nagasawa Hakuho (6.16%, 6.08%, 5.65%, 5.25%, 5.23%, and 5.20%). The content of the total polyphenol of seed extracts ranged from 18.33 to 27.08 mg/g. The antioxidant in vitro assays including the DPPH and ABTS radical scavenging activities were the highest with Hong Bak and Kawanakawase Hakuto. The tyrosinase inhibition activity greatly affected the Takinosawa Gold and Kawanakawase Hakuto. Compared to the elastase inhibition activity of 6 cultivars, Madoka was higher than others, while the lowest was Hong Bak. The significant correlation coefficient values were determined (more than p=0.05) between the yield, total polyphenol content, antioxidant activities, tyrosinase inhibition, and elastase inhibition activities. The results suggested that the total polyphenol content has a high positive value on the DPPH, ABTS radical scavenging activity, and tyrosinase inhibition activity. Also, the tyrosinase inhibition activity and elastase inhibition activity showed a significant correlation (r=+0.594). The ABTS inhibition activity and tyrosinase inhibition activity that is the most suitable character represented significant positive correlation (r=+0.846). This data suggested that the methanol extracts from these unripe peaches could be potential candidates for natural cosmetics.

Safety Evaluation of Black Garlic Extract for Development of Cosmeceutical Ingredients -Skin irritation and Sensitization Studies- (화장품 소재로서의 흑마늘 추출물에 대한 안전성 평가 -1차 피부자극 실험 및 감작성 중심으로-)

  • Lee, Hyun-Sun;Kim, Seon-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.8
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    • pp.1213-1219
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    • 2010
  • We evaluated the anti-aging potential and safety of black garlic extract for cosmeceutical ingredient. Black garlic was made by spontaneous fermentation for 40 days at $60{\sim}70^{\circ}C$, 85~95% RH without any additives. The 10% black garlic extract had sweet odor, antioxidant activities and inhibitory activities of skin againg enzymes such as tyrosinase and elastase. The skin safety was performed to evaluate of potential toxicity using the primary irritation test and skin sensitization test. The black garlic extract did not show any adverse reactions such as erythema and edema on intact skin sites at primary irritation test, but on abraded sites, some experimental animals showed very slight erythema. So, the black garlic extract was classified as a practically non-irritating material based on the score 0.23 of primary irritation index. The skin sensitization study was tested by the guinea pig maximization test (GPMT) and Freund's complete adjuvant (FCA) with intradermal injection of 10% black garlic extract. The skin sensitization test showed no skin sensitization. The allergic sensitization depends on tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and interleukin-6 (IL-6). The concentration of IL-6 on challenged tissue of treated with black garlic extract was not significantly different with negative control group (saline treated group). Based on this study, the potential for black garlic as a cosmeceutical ingredient was proven.

Effects of EGF, $\beta-ME$, Glucose, $O_2$ Concentrations and Fibroblasts Subculture on the Development of Porcine NT Embryos (EGF, $\beta-ME$, Glucose, $O_2$ 농도 및 Fibroblast Subculture가 핵이식 배의 체외발생에 미치는 영향에 관한 연구)

  • Quan J. H.;Wang A. G.;Kim S. K.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.147-156
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    • 2005
  • 본 연구는 NCSU-23과 PZM-3 배양액에 EGF, $\beta-ME$와 glucose의 첨가가 돼지 난자의 체외성숙에 미치는 영향과 배양조건을 다르게 하여 계대배양한 섬유아세포를 이용한 핵이식배를 다른 배양액과 산소조건에서 배양하였을 때 체외발생율에 미치는 영향을 조사하였다. 핵이식 배를 20ng/ml EGF를 첨가 또는 첨가하지 않은 NCSU-23 및 PZM-3 배양액에서 배양하였을 때 배반포로의 체외 발생율은 각각 $12.0\pm1.3\%,\;9.6\pm1.9\%,\;10.9\pm2.1\%,\;9.1\pm2.3\%$였다. 핵이식 배를 $25{\mu}M\;\beta-ME$를 첨가 또는 첨가하지 않은 NCSU-23 및 PZM-3 배양액에서 144시간 배양하였을 때 배반포로의 체외 발생율은 각각 $9.6\pm1.7\%,\;7.3\pm2.3\%,\;11.9\pm1.8\%$$7.4\pm2.1\%$였다. $\beta-ME$를 첨가한 PZM-3 배양액에서 배양하였을 때 배반포로의 체외 발생율은 $\beta-ME$를 첨가하지 않은 배양액에서 배양한 배보다 높은 체외발생율을 나타냈다. (p<0.05). 핵이식 배를 1.5mM glucose를 첨가 또는 첨가하지 않은 NCSU-23 및 PZM-3 배양액에서 배양하였을 때 배반포로의 체외 발생율은 각각 $9.4\pm2.2\%,\;6.8\pm2.7\%,\;10.9\pm2.4\%$$8.9\pm2.6\%$였다. Glucose를 첨가한 NCSU-23과 PZM-3 배양액에서 배양하였을 때 배반포로의 체외 발생율은 glucose를 첨가하지 않은 배양액에서 배양한 배보다 높은 체외 발생율을 나타냈다. 핵이식 배를 NCSU-23 및 PZM-3 배양액과 $5\%$$20\%$ 산소 조건에서 배양하였을 때 배반포로의 체외 발생율은 각각 $11.1\pm1.8\%,\;9.8\pm1.4\%,\;12.5\pm1.6\%$$10.9\pml.5\%$였다. NUSU-23과 PZM-3 배양액에서 $5\%$ 산소 조건에서 배양하였을 때 $20\%$산소 조건에서 배양한 난자보다 높은 체외 발생율을 나타났다. 섬유아세포를 NCSU-23 배양액에서 배양하여 공여자세포로 이용하여 10 및 $11\~15\;passage$ 이내로 계대배양하였을 때의 융합율은 $60.0\~73.3\%,\;52.5\%$였다. 섬유아세포를 PZM-3 배양액에서 배양하여 공여자세포로 이용하여 10 및 $11\~15passage$ 이내의 계대배양시의 융합율은 $60.4\~75.1\%$$58.7\%$였다.