• Title/Summary/Keyword: In vitro Development

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Assessment of In vitro Antioxidant, Antidiabetic and Cytotoxic Activities of Sphaeranthus africanus Extracts

  • Tran Thi Huyen;Julien Dujardin;Nguyen Thi Thu Huong;Chung Thi My Duyen;Nguyen Hoang Minh;Ha Quang Thanh;Dao Tran Mong;Ly Hai Trieu;Nguyen Mai Truc Tien;Mai Thanh Chung;Nguyen Nhat Minh;Nguyen Thi Ngoc Dan;Huynh Loi
    • Natural Product Sciences
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    • v.29 no.2
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    • pp.98-103
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    • 2023
  • Sphaeranthus africanus is commonly used as a traditional remedy for sore throats and pain treatment in Vietnam. The aerial parts have been studied for its anti-inflammatory and anti-proliferative properties. However, the antioxidant and antidiabetic potential of the plant has not been explored. In this work, hydrophilic extracts of the plant's aerial parts were prepared in order to investigate its antioxidant and anti-diabetic properties. Also, the cytotoxicity of the root was evaluated and compared to that of the aerial parts. All of the extracts inhibited lipid peroxidation with IC50 values ranging from 2.05 to 3.56 ㎍/mL, indicating substantial antioxidant activity. At an IC50 value of 4.80 ㎍/mL, the 50% ethanol extract exhibited the most potent inhibition of α-glucosidase. The cytotoxic activity of root extracts is 2 to 5-fold less than that of the aerial parts. Nevertheless, dichloromethane and ethyl acetate extracts of the root demonstrated a selective effect on leukemia cells, with no harm towards the normal HEK-293 cell line. This work provides a scientific support for the antioxidant and antidiabetic activity of the plant. Hence, it may find a promising material for the development of novel antioxidant and antidiabetic agents. More research can be conducted on the phytochemistry and anticancer activities of the plant's root.

Effects of Cooling Rate and Equilibration Time on the Survival and Development of Frozen-thawed Bovine Immature Oocytes (동결속도와 평형시간이 소 미성숙 난자의 동결 융해후 생존율에 미치는 영향)

  • 양병철;양보석;성환후;임기순;최선호;장원경;진동일;임경순
    • Korean Journal of Animal Reproduction
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    • v.25 no.1
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    • pp.19-28
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    • 2001
  • The present study was undertaken to investigate the effects of cooling rate and equilibration time on the survival, in vitro maturation and development to embryos of frozen-thawed bovine immature oocytes(Germinal Vesicle Stage). The cryoprotectants are used 10% ethylene glycol(EG) as permeating cryoprotectant and 0.05M soc.ose(S) or trehalose(T) as low molecular weight nonpermeating cryoprotectants and 5% ficoll(F) or polyvinylpyrrolidone(PVP) as high molecular weight nonpermeating cryoprotectants. Four freezing solution were uysed in this experiment(EFT: 10% EG + 5% F + 0.05M T, EFS: 10% EG + 5% F + 0.05M S, EPT: 10% EG + 5% P + 0.05M T, EPS: 10% EG + 5% P + 0.05M S). The best equilibration time and freezing solution was 15 min in EPT(83% survival rate of frozen-thawed bovine immature oocytes). When frozen-thawed bovine oocytes were cultured following IVM and IVF, there was no significant difference in cleavage and development rates among the EFT, EFS, EPT and EPS solutions. When 9 blastocysts derived from frozen bovine oocytes were transferred to 6 recipients, two recipients were pregnant. And one was aborted at 45 days of pregnancy and the other had a stillbirth.

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Matrix metalloproteinase-1 suppression and type-1 procollagen synthesis promoting effects of Uncaria gambir (아선약 추출물의 matrix metalloproteinase-1(MMP-1) 억제 및 제1형 프로콜라겐 합성 촉진 활성)

  • Lee, Jaemin;Kim, Dong-Hee;Kim, Eun-Woo;Kwon, Hyun Ju;Kim, Byung Woo;Cho, Jae-Hyon;Kim, Hyun-Joo;Kim, Tae Hoon
    • Food Science and Preservation
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    • v.24 no.1
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    • pp.93-99
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    • 2017
  • Collagen synthesis is decreased and matrix metalloproteinase-1 (MMP-1) levels are increased in naturally aged human skin, and these alterations cause changes such as skin wrinkling and decreased elasticity. As a part of our ongoing search for bioactive ingredients, MMP-1 inhibitory and type-1 procollagen synthesis inducing activities of aqueous methanolic extract of manufactured gambir product from Uncaria gambir were investigated in in vitro bioassay systems. In addition, total phenolic contents were quantified using a spectrophotometric method. Among tested samples, 40% MeOH eluate from 80% methanolic extract of manufactured U. gambir using open column chromatography packed with Diaion HP-20 resin showed significant MMP-1 inhibitory activities with an $IC_{50}$ value of $15.6{\pm}1.3{\mu}g/mL$. Furthermore, type-1 procollagen synthesis promoting property of 40% MeOH eluate ($IC_{50}$ value; $6.9{\pm}0.7{\mu}g/mL$) from 80% methanolic extract of manufactured gambir was higher than other eluates. Additionally, the present investigation revealed that 40% MeOH eluate of manufactured gambir product contained a high level of total phenolic compounds. The result suggests a distinct relationship between anti-wrinkle activity and total phenolic contents, and manufactured gambir product could be considered a new effective source of natural bioactive ingredients. Systematic investigation of manufactured gambir product will be performed for further development of its biological properties.

Effects of the Extracts from Gyrophora esculenta and Coriolus versicolor judae Mycelia on the Growth of Intestinal Bacteria (석이버섯과 운지버섯 균사체 추출물이 장내 세균의 생육에 미치는 영향)

  • Park, Kyoung-Ran;Lee, Woon-Jong;Cho, Min-Gyu;Park, Eui-Seok;Jeong, Jun-Young;Kwon, Oh-Sung;Yoon, Hyang-Sik;Kim, Kwang-Yup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.6
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    • pp.820-825
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    • 2010
  • The functional food components from various Basidiomycota were investigated to improve human intestinal microflora, especially associated with obesity. EtOH extract from Gyrophora esculenta fruit body and Coriolus versicolor judae mycelia showed antimicrobial activities on Eubacterium limosum, Clostridium perfrigens, Clostridium paraputrificum, Clostridium difficile and Clostridium ramosum, and on Bacteroides fragilis, respectively. Although the 80% EtOH extract from G. esculenta fruit body and hot-water extract from C. versicolor judae mycelia did not reduce weight of the rats in the high fat diet, these extracts showed stability at high temperatures and at wide pH ranges. In the rat group of feeding 80% EtOH extract from G. esculanta fruit body, Bifidobacterium spp. were increased and Clostridium spp. and Eubacterium spp. were decreased compared to the high fat feeding group. Also sensory evaluation was carried out for the development of prototype drink product. These results demonstrated the possibilities of C. versicolor judae and G. esculenta as a functional food components to control intestinal microbial flora.

Isoflavone Composition and Estrogenic Activity of Germinated Soybeans (Glycine max) according to Variety (품종별 발아 콩(Glycine max)의 아이소플라본 조성 및 In Vitro 에스트로겐 유사활성)

  • Kim, Min Young;Jang, Gwi Yeong;Ji, Yeong Mi;Kim, Kyung Mi;Kim, Hongsik;Lee, Junsoo;Jeong, Heon Sang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.10
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    • pp.1430-1437
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    • 2016
  • This study was performed to investigate the changes in isoflavone composition, estrogenic activity and antiestrogenic activity of soybean cultivars of different isoflavone content (Aga 8, Uram, Cheongja 3, and Dawon) with germination. Total isoflavone contents of Aga 8, Dawon and Cheongja 3 was increased from 2,671.74, 261.08 and 2,240.08 to 2,977.50, 966.13 and $2,354.11{\mu}g/g$, respectively after germination except for Uram cultivars, and highest contents of total isoflavone showed $2,977.50{\mu}g/g$ and $2,354.11{\mu}g/g$, respectively in Aga 8 and Cheongja 3 after germination. MTT cell proliferation assay using MCF-7 cells revealed that germinated soybean of Aga 8 and Cheonja 3 obtained not only contained a high content of isoflavone but also had estrogenic activity. Estrogenic activity of Aga 8 and Cheongja 3 soybean extracts increased from 116.21% and 101.60% to 135.34% and 121.05% after germination. These results suggest that germinated soybean of Aga 8 and Cheongja 3 might have a potential preventive effect on estrogen-deficient diseases.

Aggregation of Human Eyelid Adipose-derived Stem Cells by Human Body Fluids

  • Song, Yeonhwa;Yun, Sujin;Yang, Hye Jin;Yoon, A Young;Kim, Haekwon
    • Development and Reproduction
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    • v.16 no.4
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    • pp.339-351
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    • 2012
  • Fetal bovine serum (FBS) is the most frequently used serum for the cultivation of mammalian cells. However, since animal-derived materials might not be appropriate due to safety issues, allogeneic human serum (HS) has been used to replace FBS, particularly for the culture of human cells. While there has been a debate about the advantages of HS, its precise effect on human adult stem cells have not been clarified. The present study aimed to investigate the effect of HS on the human eyelid adipose stem cells (HEACs) in vitro. When HEACs were cultivated in a medium containing 10% HS, many cells moved into several spots and aggregated there. The phenomenon was observed as early as 9 days following 10% HS treatment, and 12 days following 5% HS plus 5% FBS treatment. However, the aggregation was never observed when the same cells were cultivated with 10% FBS or bovine serum albumin. To examine whether cell density might affect the aggregation, cells were seeded with different densities on 12-well dish. Until the beginning of aggregation, cells seeded at low densities exhibited the longest culture period of 16 days whereas cells seeded at high densities showed the shortest period of 9 days to form aggregation. The number of cells was $15.1{\pm}0.2{\times}10^4$ as the least for the low density group, and $29.3{\pm}2.8{\times}10^4$ as the greatest for the high density group. When human cord blood serum or normal bovine serum was examined for the same effect on HEACs, interestingly, cord blood serum induced the aggregation of cells whereas bovine serum treatment has never induced. When cells were cultivated with 10% HS for 9 days, they were obtained and analyzed by RT-PCR. Compared to FBS-cultivated HEACs, HS-cultivated HEACs did not express VIM, and less expressed GATA4, PALLD. On the other hand, HS-cultivated HEACs expressed MAP2 more than FBS-cultivated HEACs. In conclusion, human adult stem cells could move and form aggregates by the treatment with human body fluids.

Antioxidant Activity of Ripe Fruit Components of Rubus coreanus: Extraction Using Porous Polymer Resins (다공성 합성흡착제를 이용한 복분자 딸기(Rubus coreanus) 농축액의 항산화 효과)

  • Choi, Se-Jin;Lee, Yeon-Sil;Kim, Jin-Kyu;Chung, Cha-Kwon;Kang, Il-Jun;Lim, Soon-Sung
    • Korean Journal of Food Science and Technology
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    • v.43 no.2
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    • pp.149-155
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    • 2011
  • This study was carried out to develop a method of removing unnecessary sugars and high-molecular weight soluble components from water extract by using various highly porous polymer resins: these resins are widely used as adsorbents for polyphenolic compounds. Each anthocyanin-rich fraction (ARF) that was eluted from column packed with 4 different resins (SEPABEADS SP207, 700, 850 and Diaion HP 20) was obtained from an aqueous extract of the ripe fruits of Rubus coreanus, which is well-known in Korea as "Bok-bun-ja". Among the above mentioned resins, Diaion HP20 had the highest efficacy and provided maximum yield of the ARF. The ARF eluted from the column packed with HP 20 exhibited the strongest antioxidant activity in vitro. Bioactive phytochemicals extracted from Rubus coreanus were investigated using the on-line HPLC-$ABTS^{{\cdot}+}$ system, and the resulting 6 peaks were identified as radical-scavenging components. By using liquid chromatography-mass spectrometry, 5 peaks were obtained, and these were identified as cyanidin-3-sambubioside, cyanidin-3-glucoside, cyanidin-3-xylosylrutinoside, cyanidin-3-rutinoside and pelargonidin-3-rutinoside. These results indicate that the use of appropriate porous resin (Diaion HP 20) leads to an increase in the yield of bioactive components and enhancement of their biological properties.

Production of Chimeric Mice Following Transgenesis of Multipotent Spermatogonial Stem Cells (유전자변형 다분화능 정원줄기세포를 이용한 키메라 생쥐의 생산)

  • Lim, Jung-Eun;Eum, Jin-Hee;Kim, Hyung-Joon;Park, Jae-Kyun;Lee, Hyun-Jung;Lee, Dong-Ryul
    • Development and Reproduction
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    • v.13 no.4
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    • pp.305-312
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    • 2009
  • Multipotent spermatogonial stem cells (mSSCs), derived from uni-potent SSC, are a type of reprogrammed cells with similar characteristics to embryonic stem cells (ESCs). The aim of this study was to evaluate the potential for transgenesis of mSSC derived from outbred mice and the production of transgenic animal by the mSSC-insertion into embryo. mSSCs, established from outbred mice (ICR strain) in the previous study, were maintained and then transfected with a lenti-viral vector expressing green fluorescent protein (GFP), CS-CDF-CG-PRE. Embryonic stem cells (ESCs) were derived from inbred transgenic mice (C57BL/6-Tg (CAG-EGFP)) and were used as an experimental control. Transfected mSSCs were well proliferated in vitro and maintained their characteristics and normal karyotype. Ten to twelve mSSCs and ESCs were collected and inserted into perivitelline space of 8-cell mouse embryos, and then transferred them into uteri of poster mothers after an additional 2-days of culture. Percentage of mSSC-derived offsprings was 4.8% (47/980) and which was lower than those (11.7% (67/572)) of ESC-derived ones (P<0.05). However, even though different genetic background of mSSC and ESC origin, the production efficiency of coat-colored chimeric offspring in mSSC group was not different when compared it with ESC (6.4% (3/47) vs. 7.5% (5/67)). From these results, we confirmed that mSSC derived from outbred mice has a pluripotency and a potential to produce chimeric embryos or mice when reaggregatation with mSSC is performed.

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Inhibitory Efficacy of Black Tea Water Extract on Melanogenesis in Melan-a Cells and Its Action Mechanisms (멜라닌 세포에서 홍차 열수추출물의 멜라닌 합성 저해능과 작용기전)

  • Choi, So-Young;Kim, Young-Chul;Chang, Byung-Soo
    • Applied Microscopy
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    • v.41 no.3
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    • pp.169-177
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    • 2011
  • To evaluate the whitening effect of Camellia sinensis water extract (CSWE), CSWE was treated to melan-a cells. Total polyphenol contents and flavonoid contents of CSWE were 102 mg/g and 87 mg/g, respectively. The electron-donating ability of CSWE revealed a dose-dependent response, showing the excellent ability of 82% at 800 ${\mu}g$/mL, and which was higher than the arbutin (48%). The CSWE significantly (p<0.001) suppressed the melanin synthesis and the development of melanocyte dendrites was inhibited in a dose-dependent manner. The CSWE significantly (p<0.001) inhibited both intra-cellular and cell-extracted tyrosinase activities. And inhibitory efficacies of CSWE on both melanin synthesis and tyrosinase activity were significantly (p<0.001) higher than the arbutin. The tyrosinase protein expression was not influenced by arbutin treatment. However, CSWE treatment significantly (p<0.001) reduced it. Both arbutin and CSWE treatment did not influence on mRNA expressions of tyrosinase, tyrosinase-related protein-1 and tyrosinase related protein-2.

The Various biofunctional effects(anticarcinogenic, antioxidative and lypolytic activity) of Pohang buchu (포항 부추의 생화학적 기능성 효과)

  • Hwang, Cher-Won;Shin, Hyun-Kil;Do, Myoung-Sool;Kim, Yun-Joo;Park, Jong-Heum;Choi, Young-Sub;Joo, Woo-Hong
    • Korean Journal of Food Science and Technology
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    • v.33 no.3
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    • pp.279-281
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    • 2001
  • Leek(Allium tuberosum Rotter) is a korean traditional herb of which leaves are used for daily food. In order to improve the potency of leek as a functional food, we investigated the several biochemical functions in vitro. The quinone reductase activity, an anticarcinogenic enzyme, was induced 2 times at the concentration of 1mg/ml of leek as compared to that of negative control(80% methanol). The induction of SOD(Superoxide dismutase) activity was also demonstrated. But the functional activities of leek were shown to be a little difference depending on the place where the leek was produced. The difference might to be due to the different environmental factors where affects the growth of leek. In an experiment to test the lypolytic activity of leek, the glycerol release was significantly increased(about 3 times to that of negative control). This was suggested that leek can be considered as a potential food for diet. The above results provide the potential food, and more profound research is requested.

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