• 제목/요약/키워드: In Vitro Development

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Adventitious Shoot Formation and Plant Regeneration from Explants of Solanum nigrum L. (까마중(Solanum nigrum L.)의 유식물 절편체에서 부정아 형성 및 식물체 재분화)

  • Oh, Soon-Ja;Koh, Seok-Chan
    • Korean Journal of Plant Resources
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    • v.25 no.2
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    • pp.277-284
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    • 2012
  • In the present study, the effects of plant growth regulators on adventitious shoot and root formation of various explants of $in$ $vitro$ seedlings of $Solanum$ $nigrum$ L. were investigated to determine the optimum conditions for the high-efficiency plant regeneration of this species. The formation of adventitious shoots was higher in leaf explants than in cotyledon, hypocotyl, or epicotyl explants at low concentrations (0.5~2.0 mg $L^{-1}$ ) of 6-benzylaminopurine (BAP). The number of adventitious shoots and the shoot length were also higher in both leaf and cotyledon explants. In particular, 2.0 mg $L^{-1}$ BAP was most effective for stimulating the induction and multiplication of adventitious shoots. In terms of root formation and root development from shoots that were separated from multiple shoots, indole butyric acid (IBA) and indole acetic acid (IAA) were more effective than ${\alpha}$-naphthalene acetic acid (NAA). The percentage of rooting as well as the number of roots per shoot (4.0), root length (7.82 cm), and shoot length (8.76 cm) was highest on MS media supplemented with 0.05 mg $L^{-1}$ IAA. Furthermore, 100% of the regenerated plantlets survived when transplanted to compost soil. These results suggest that leaf explants are the best source for the high-efficiency regeneration of $S.$ $nigrum$ L., and that 2.0 mg $L^{-1}$ BAP and 0.05 mg $L^{-1}$ IAA are the best conditions for shoot and root induction, respectively.

Studies on the Survival Rates after Ultrarapidly Frozen-Thawing of Porcine Embryos (돼지 수정난의 초급속 동결 융해후의 생존성에 관한 연구)

  • 김상근;이봉구
    • Korean Journal of Animal Reproduction
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    • v.16 no.2
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    • pp.125-131
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    • 1992
  • This study was carried out to investigate the effects of concentration and equilibration time of cryoprotective agents on the survival rate of slowly and ultrarapidly frozen porcine embryos. The porcine embryos following dehydration by cryoprotective agents and a various concentration of sucrose were directly plunged into liquid nitrogen and thawed in 38$^{\circ}C$ water bath. Survival rate was defined as development rat to the morula and blastocyst stage after in vitro culture or by FDA test. The results are summarized as follows : 1. The survival rates of porcine embryos after ultrarapid frozen-thawing in the freezing medium of 0.25M sucrose added 2.0, 2.5, 3.0, 3.5, or 4.0M glycerol was 65.3, 61.8, 64.3, 59.4 or 39.4%, respectively. Addition of 0.25M sucrose into the freezing medium containing 2.0M glycerol showed higher survival rate than those of 2.5~4.0M glycerol. 2. The survival rates of porcine embryos after ultraradpid frozen-thawing in the freezing medium of 0.25M sucroese added 2.0, 2.5, 3.0, 3.5 or 4.0M DMSO was 65.6, 67.6, 68.6, 60.6 or 23.6%, respectively. However, addition of 0.25M sucrose into the freezing medium containing 3.0M DMSO showed higher survival rate than those of 2.0, 2.5, 3.5 or 4.0M DMSO. 3. The survival rates of porcine embryos after ultrapid frozen-thawing in the freezing medium of 0.25M sucrose added 2.0, 2.5, 3.0, 3.5 or 4.0M propanediol was 63.2, 60.3, 62.1, 52.3 or 24.3%, respectively. Addition of 0.25M sucroese into the freezing medium containing 2.0M propanediol showed higher survival rate than those of 2.5~4.0M glycerol. 4. The survival rates of porcine embryos after ultrarapid frozen-thawing the freezing medium of 2.0M glycerol added 0.10, 0.25, 0.50 or 0.75M sucrose was 61.8, 70.8, 67.6 or 52.2%, respectively. Addition of 2.0M glycerol into the freezing medium containing 0.25M sucreose showed higher survival rate than that those of 0.10, 0.50 or 0.75M sucrose. 5. The higher suvival rate of porcine embryos were attained at short period of equilibration time 92.5~5min.) in the freezing medium added 0.25M sucreose and 3.0M compared to those of 10 or 20min. equilibration time in the same condition.

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Antioxidant Activity and Cytotoxicity against Human Cancer Cells of Glycyrrhiza New Varieties : A Comparison with Glycyrrhiza Official Compendia (감초 신품종과 약전 수재 감초 추출물의 항산화 활성 및 암세포 독성 비교 연구)

  • Kim, Minhee;Kang, Myunghoon;Lee, Jeonghoon;Leem, Kang-Hyun;An, Hyo-Jin;Jin, Jong-Sik;Lee, Jong-Hyun;Chang, Jaeki;Seong, Shin;Kim, Wonnam
    • The Korea Journal of Herbology
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    • v.36 no.3
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    • pp.15-24
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    • 2021
  • Objectives : The Glycyrrhiza new varieties, WONGAM and SINWONGAM, were developed through interspecific cross between Glycyrrhiza glabra and Glycyrrhiza uralensis by the National Institute of Horticultural and Herbal Science, Rural Development Administration in Korea. This in vitro study was undertaken to compare the antioxidant and cytotoxic effects between Glycyrrhiza new varieties (WONGAM and SINWONGAM) and official compendia (Glycyrrhiza glabra and Glycyrrhiza uralensis). Methods : Antioxidant activity was determined by DPPH (1,1-diphenyl-2-picrylhy drazyl), ABTS (2,2-azino-bis (3-rthylbenz-thiazoline-6-sulfonic acid)) diammonium salt, Nitrite radical scavenging assay, and Reducing Power assay. Cytotoxicity was determined by MTT assay and cell morphology was observed by an inverted microscope. Results : The DPPH, ABTS, Nitrite radical scavenging activities and reducing power of Glycyrrhiza glabra, Glycyrrhiza uralensis, WONGAM, and SINWONGAM were evaluated at different concentrations (0, 10, 50, 100, 500, 1000 ㎍/㎖). Glycyrrhiza glabra, Glycyrrhiza uralensis, WONGAM, and SINWONGAM showed similar dose-dependent increase in antioxidant activities. The cytotoxic effects with increasing doses of Glycyrrhiza new varieties and official compendia did not differ in HCT116, HT29, A549, MDA-MB231, PC3, ACHN, and HeLa cells. However, significant difference in cytotoxicity were observed in AGS, MCF7 and Hep3B cells by Glycyrrhiza glabra, Glycyrrhiza uralensis, WONGAM, and SINWONGAM. Conclusions : These results showed that Glycyrrhiza new varieties and official compendia acts as a potent antioxidant. Also, the finding that equivalent cytotoxic potency was observed in a cell dependent manner. Our study suggests that Glycyrrhiza new varieties may offer a wide-variety of health benefits.

Yeosin-san Increases Female Fertility through Inducing Uterine Receptivity and Ovarian Function

  • Choi, Hee Jung;Joo, Bo Sun;Park, Mi Ju;Park, Min Jung;Bae, Boram;Kim, Bo Sung;Park, Hye Rin;Kim, Keuk Jun;Yang, Hee Jin;Yoo, Jeong Eun;Chung, Tae Wook;Joo, Jongkil;Ha, Ki Tae
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.33 no.2
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    • pp.141-150
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    • 2019
  • Despite the development of assisted reproduction technologies (ART) including in vitro fertilization (IVF), the poor ovarian response and endometrial receptivity remains clinically a major unmet need. Although these problems are difficulties to solve in infertility treatment, there are no good therapeutic option yet. Traditional herbal remedies and acupuncture, therefore are being proposed as alternative treatment. Our group found that traditional herbal medicines such as Paeonia lactiflora L.(PL, 芍藥), Cyperus rotundus L.(CR, 香附子), and Perilla frutescens (PF, 紫蘇葉) could improve endometrial receptivity. In this study, we found out Yeosin-san (如神散) as an optimal herbal formula via combination of the previously established herbal medicines. Yeosin-san is a traditional Korean medical formula which was established by Ziming Jin (陳自明) and recorded in Furendaiquanliangfang (婦人大全良方) at first. The formula traditionally used for treating abnormal uterine bleeding and leukorrhea. It showed a highest effect on leukemia inhibitory factor (LIF) expression and on the adhesion between trophoblastic cells and endometrial cells. In addition, it has been shown that the Yeosin-san not only increases the endometrial receptivity to improve the embryo implantation but also enhances the ovary function by expressing the angiogenesis-related genes. Here we suggest that Yeosin-san could be a novel and effective candidate for treating female infertility.

What Is Cultured Meat? (배양육이란 무엇인가?)

  • Huh, Man Kyu
    • Journal of Life Science
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    • v.31 no.6
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    • pp.587-594
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    • 2021
  • By 2050, 70% more food will be needed to fulfill the demands of a growing population. Among the solutions, cultured meat or clean meat is presented as a sustainable alternative for consumers. Scientists have begun to leverage knowledge and tools accumulated in the fields of stem cell and tissue engineering in efforts aimed at the development of cell-based meat. Cultured meat has to recreate the complex structure of livestock muscles with a few cells. Cells start to divide after they are cultured in a culture medium, which provides nutrients, hormones, and growth factors. An initial problem with this type of culture is the serum used, as in vitro meat aims to be slaughter free. Thus, it is contradictory to use a medium made from the blood of dead calves. The serum is expensive and affects to a large extent the production cost of the meat. A positive aspect related to the safety of cultured meat is that it is not produced from animals raised in confined spaces and slaughtered in inhumane conditions. Thus, the risk of an outbreak is eliminated, and there is no need for vaccinations and animal welfare issues. The production of cultured meat is presented as environmentally friendly, as it is supposed to produce less greenhouse gas, consume less water, and use less land in comparison to conventional meat production.

Effects of exploration and molecular mechanism of CsV on eNOS and vascular endothelial functions

  • Zuo, Deyu;Jiang, Heng;Yi, Shixiong;Fu, Yang;Xie, Lei;Peng, Qifeng;Liu, Pei;Zhou, Jie;Li, Xunjia
    • Advances in nano research
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    • v.12 no.5
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    • pp.501-514
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    • 2022
  • This study aimed to investigate the effects and potential mechanisms of Chikusetsusaponin V (CsV) on endothelial nitric oxide synthase (eNOS) and vascular endothelial cell functions. Different concentrations of CsV were added to animal models, bovine aorta endothelial cells (BAECs) and human umbilical vein endothelial cells (HUVECs) cultured in vitro. qPCR, Western blotting (WB), and B ultrasound were performed to explore the effects of CsV on mouse endothelial cell functions, vascular stiffness and cellular eNOS mRNA, protein expression and NO release. Bioinformatics analysis, network pharmacology, molecular docking and protein mass spectrometry analysis were conducted to jointly predict the upstream transcription factors of eNOS. Furthermore, pulldown and ChIP and dual luciferase assays were employed for subsequent verification. At the presence or absence of CsV stimulation, either overexpression or knockdown of purine rich element binding protein A (PURA) was conducted, and PCR assay was employed to detect PURA and eNOS mRNA expressions, Western blot was used to detect PURA and eNOS protein expressions, cell NO release and serum NO levels. Tube formation experiment was conducted to detect the tube forming capability of HUVECs cells. The animal vasodilation function test detected the vasodilation functions. Ultrasonic detection was performed to determine the mouse aortic arch pulse wave velocity to identify aortic stiffness. CsV stimulus on bovine aortic cells revealed that CsV could upregulate eNOS protein levels in vascular endothelial cells in a concentration and time dependent manner. The expression levels of eNOS mRNA and phosphorylation sites Ser1177, Ser633 and Thr495 increased significantly after CsV stimulation. Meanwhile, CsV could also enhance the tube forming capability of HUVECs cells. Following the mice were gavaged using CsV, the eNOS protein level of mouse aortic endothelial cells was upregulated in a concentration- and time-dependent manner, and serum NO release and vasodilation ability were simultaneously elevated whereas arterial stiffness was alleviated. The pulldown, ChIP and dual luciferase assays demonstrated that PURA could bind to the eNOS promoter and facilitate the transcription of eNOS. Under the conditions of presence or absence of CsV stimulation, overexpression or knockdown of PURA indicated that the effect of CsV on vascular endothelial function and eNOS was weakened following PURA gene silence, whereas overexpression of PURA gene could enhance the effect of CsV upregulating eNOS expression. CsV could promote NO release from endothelial cells by upregulating the expression of PURA/eNOS pathway, improve endothelial cell functions, enhance vasodilation capability, and alleviate vessel stiffness. The present study plays a role in offering a theoretical basis for the development and application of CsV in vascular function improvement, and it also provides a more comprehensive understanding of the pharmacodynamics of CsV.

Quantitation of relationship and development of nutrient prediction with vibrational molecular structure spectral profiles of feedstocks and co-products from canola bio-oil processing

  • Alessandra M.R.C.B. de Oliveira;Peiqiang Yu
    • Animal Bioscience
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    • v.36 no.3
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    • pp.451-460
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    • 2023
  • Objective: This program aimed to reveal the association of feed intrinsic molecular structure with nutrient supply to animals from canola feedstocks and co-products from bio-oil processing. The special objective of this study was to quantify the relationship between molecular spectral feature and nutrient availability and develop nutrient prediction equation with vibrational molecular structure spectral profiles. Methods: The samples of feedstock (canola oil seeds) and co-products (meals and pellets) from different bio-oil processing plants in Canada (CA) and China (CH) were submitted to this molecular spectroscopic technique and their protein and carbohydrate related molecular spectral features were associated with the nutritional results obtained through the conventional methods of analyses for chemical and nutrient profiles, rumen degradable and intestinal digestible parameters. Results: The results showed that the spectral structural carbohydrates spectral peak area (ca. 1,487.8 to 1,190.8 cm-1) was the carbohydrate structure that was most significant when related to various carbohydrate parameters of canola meals (p<0.05, r>0.50). And spectral total carbohydrate area (ca. 1,198.5 to 934.3 cm-1) was most significant when studying the various carbohydrate parameters of canola seeds (p<0.05, r>0.50). The spectral amide structures (ca. 1,721.2 to 1,480.1 cm-1) were related to a few chemical and nutrient profiles, Cornell Net Carbohydrate and Protein System (CNCPS) fractions, truly absorbable nutrient supply based on the Dutch protein system (DVE/OEB), and NRC systems, and intestinal in vitro protein-related parameters in co-products (canola meals). Besides the spectral amide structures, α-helix height (ca. 1,650.8 to 1,643.1 cm-1) and β-sheet height (ca. 1,633.4 to 1,625.7 cm-1), and the ratio between them have shown to be related to many protein-related parameters in feedstock (canola oil seeds). Multi-regression analysis resulted in moderate to high R2 values for some protein related equations for feedstock (canola seeds). Protein related equations for canola meals and carbohydrate related equations for canola meals and seeds resulted in weak R2 and low p values (p<0.05). Conclusion: In conclusion, the attenuated total reflectance Fourier transform infrared spectroscopy vibrational molecular spectroscopy can be a useful resource to predict carbohydrate and protein-relates nutritional aspects of canola seeds and meals.

Comparison of Regeneration Conditions in Seven Pepper (Capsicum annuum L.) Varieties (7종의 고추(Capsicum annuum L.) 재분화 조건 비교)

  • Min-Su Kim;Yun-Jeong Han;Sharanya Tripathi;Jinwoo Kwak;Jin-Kyung Kwon;Byoung-Cheorl Kang;Jeong-Il Kim
    • Korean Journal of Plant Resources
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    • v.36 no.5
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    • pp.527-539
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    • 2023
  • Pepper (Capsicum annuum L.) is an important vegetable and spice crop that has been cultivated worldwide. Pepper fruits have unique taste and aroma, providing a variety of antioxidants and compounds important for human health, which makes a high economic value. In addition, there is a high demand for new pepper varieties, according to consumer's preference. However, pepper is a recalcitrant plant for in vitro tissue and organ differentiation and plant regeneration, which makes it difficult to develop demanded varieties using newly developed technologies such as genetic engineering and gene editing. In this study, tissue culture and regeneration conditions were investigated using seven pepper varieties that were obtained from the core-collection of Seoul National University. We observed callus and bud induction and shoot formation using several media composition composed of different cytokinins and auxin concentrations. As a result, it was found that there were differences in callus induction and shoot formation of each variety depending on the hormone composition, and the highest regeneration was shown when the medium containing Zeatin Riboside and the petioles of seedlings were used. In particular, out of seven pepper varieties, CMV980 exhibited a higher regeneration efficiency (approximately 48%) than other varieties, followed by Yuwolcho. Therefore, this study provides CMV980 and Yuwolcho as good candidates that can be used for pepper transformation, which might contribute to the development of various varieties through gene editing technology in the future.

Cigarette Smoke Extract-Treated Mouse Airway Epithelial Cells-Derived Exosomal LncRNA MEG3 Promotes M1 Macrophage Polarization and Pyroptosis in Chronic Obstructive Pulmonary Disease by Upregulating TREM-1 via m6A Methylation

  • Lijing Wang;Qiao Yu;Jian Xiao;Qiong Chen;Min Fang;Hongjun Zhao
    • IMMUNE NETWORK
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    • v.24 no.2
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    • pp.3.1-3.23
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    • 2024
  • Cigarette smoke extract (CSE)-treated mouse airway epithelial cells (MAECs)-derived exosomes accelerate the progression of chronic obstructive pulmonary disease (COPD) by upregulating triggering receptor expressed on myeloid cells 1 (TREM-1); however, the specific mechanism remains unclear. We aimed to explore the potential mechanisms of CSE-treated MAECs-derived exosomes on M1 macrophage polarization and pyroptosis in COPD. In vitro, exosomes were extracted from CSE-treated MAECs, followed by co-culture with macrophages. In vivo, mice exposed to cigarette smoke (CS) to induce COPD, followed by injection or/and intranasal instillation with oe-TREM-1 lentivirus. Lung function and pathological changes were evaluated. CD68+ cell number and the levels of iNOS, TNF-α, IL-1β (M1 macrophage marker), and pyroptosis-related proteins (NOD-like receptor family pyrin domain containing 3, apoptosis-associated speck-like protein containing a caspase-1 recruitment domain, caspase-1, cleaved-caspase-1, gasdermin D [GSDMD], and GSDMD-N) were examined. The expression of maternally expressed gene 3 (MEG3), spleen focus forming virus proviral integration oncogene (SPI1), methyltransferase 3 (METTL3), and TREM-1 was detected and the binding relationships among them were verified. MEG3 increased N6-methyladenosine methylation of TREM-1 by recruiting SPI1 to activate METTL3. Overexpression of TREM-1 or METTL3 negated the alleviative effects of MEG3 inhibition on M1 polarization and pyroptosis. In mice exposed to CS, EXO-CSE further aggravated lung injury, M1 polarization, and pyroptosis, which were reversed by MEG3 inhibition. TREM-1 overexpression negated the palliative effects of MEG3 inhibition on COPD mouse lung injury. Collectively, CSE-treated MAECs-derived exosomal long non-coding RNA MEG3 may expedite M1 macrophage polarization and pyroptosis in COPD via the SPI1/METTL3/TREM-1 axis.

Increased mRNA Related Ovarian Maturation during Induction of Maturational Competence in Red Seabream, Pagrus major (참돔, Pagrus major의 성숙능력 유도시 증가된 난성숙 관련 mRNA)

  • Choi, Cheol-Young;Chang, Young-Jin;Takashima, Fumio
    • Development and Reproduction
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    • v.4 no.1
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    • pp.125-131
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    • 2000
  • This study has used differential display-PCR, to amplify genes transcribed during the ovarian maturation induced by human chorionic gonadotropin (hCG). The cDNA expressed at the times of acquisition of oocyte maturational competence in red seabream (Pagrus major) following treatment with hCG was amplified and cloned. A full-length of cDNA for p. major was isolated using differential display-PCR and 5'RACE. This cDNA clone contained 2,662 nucleotides including the open reading frame that encoded 434 amino acids. Homology analyses, using the GenBank and EMBL general database searches, indicated that the nucleotides sequence of the cDNA does not have high homology with any other genes. This cDNA was judged to be a gene, which induction of maturational competence coincides with increase of mRNA related ovarian maturation. Consensus sequences which were consistent with protein kinase C phosphorylation sites and casein kinase II phosphorylation sites were identified. in vitro, the transcription level of mRNA related ovarian maturation increased between 9hr and 24hr following treatment of ovarian follicles with hCG. It was also increased after GtH-II (300 ng/ml) stimulation. Furthermore, in vivo, mRNA related ovarian maturation was rarely expressed prior to the acquisition of oocyte maturational competence, but was strongly expressed after the acquisition of oocyte maturational competence, suggesting that the hCG induction of maturational competence is brought about by the de novo synthesis of the mRNA related ovarian maturation in p. major.

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