• Title/Summary/Keyword: In Vivo and In Vitro Function

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Establishing porcine jejunum-derived intestinal organoids to study the function of intestinal epithelium as an alternative for animal testing

  • Bo Ram Lee;Sun A Ock;Mi Ryung Park;Min Gook Lee;Sung June Byun
    • Journal of Animal Reproduction and Biotechnology
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    • v.39 no.1
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    • pp.2-11
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    • 2024
  • Background: The small intestine plays a crucial role in animals in maintaining homeostasis as well as a series of physiological events such as nutrient uptake and immune function to improve productivity. Research on intestinal organoids has recently garnered interest, aiming to study various functions of the intestinal epithelium as a potential alternative to an in vivo system. These technologies have created new possibilities and opportunities for substituting animals for testing with an in vitro model. Methods: Here, we report the establishment and characterisation of intestinal organoids derived from jejunum tissues of adult pigs. Intestinal crypts, including intestinal stem cells from the jejunum tissue of adult pigs (10 months old), were sequentially isolated and cultivated over several passages without losing their proliferation and differentiation using the scaffold-based and three-dimensional method, which indicated the recapitulating capacity. Results: Porcine jejunum-derived intestinal organoids showed the specific expression of several genes related to intestinal stem cells and the epithelium. Furthermore, they showed high permeability when exposed to FITC-dextran 4 kDa, representing a barrier function similar to that of in vivo tissues. Collectively, these results demonstrate the efficient cultivation and characteristics of porcine jejunum-derived intestinal organoids. Conclusions: In this study, using a 3D culture system, we successfully established porcine jejunum-derived intestinal organoids. They show potential for various applications, such as for nutrient absorption as an in vitro model of the intestinal epithelium fused with organ-on-a-chip technology to improve productivity in animal biotechnology in future studies.

Unexpected Estrogenicity of Endocrine Disruptors may Evoke a Failure of Pregnancy Derived from Uterine Function: Overview of Their Possible Mechanism(s) through Steroid Receptors

  • Dang, V.H.;Nguyen, T.H.;Choi, K.C.;Jeung, E.B.
    • Journal of Embryo Transfer
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    • v.22 no.4
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    • pp.199-208
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    • 2007
  • Although many studies have focused on the biological and toxicological effects of phenol products, in particular, in reproductive tracts, the data about their effects in this estrogenic responsive tissue are much less clear. In addition, the in vitro and in vivo data concerning ED-adverse impacts in other endocrine organs, i.e. pituitary gland, are not understood well either. Thus, a further study is needed for providing a new insight into possible impacts of estrogenic EDs including phenol products in humans and wildlife. A combination of in vitro and in vivo system for examining EDs may bring better understanding into the regulatory mechanisms underlying EDs-induced events. In addition, this information may support for developing optimal screening methods of estrogenic EDs, in particular, phenol products.

Immunostimulatory Effect of Fermented Red Ginseng in the Mouse Model

  • Park, Sang-Yong;Kim, Ho-Bin;Kim, Jeong-Hoon;Lee, Joo-Mi;Kim, Sang-Rae;Shin, Heon-Sub;Yi, Tae-Hoo
    • Preventive Nutrition and Food Science
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    • v.19 no.1
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    • pp.10-18
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    • 2014
  • In this study, Woongjin fermented red ginseng extract (WFRG) was evaluated for its potential ability to act as an adjuvant for the immune response of mice. For the in vitro study, macrophages were treated with serial concentrations ($1{\mu}g/mL$, $10{\mu}g/mL$, and $100{\mu}g/mL$) of WFRG. For in vivo studies, mice were administered different concentrations (10 mg/kg/day, 100 mg/kg/day, and 200 mg/kg/day) of WFRG orally for 21 days. In vitro, the production of nitric oxide and TNF-${\alpha}$ by RAW 264.7 cells increased in a dose-dependent manner. In vivo, WFRG enhanced the proliferation of splenocytes induced by two mitogens (i.e., concanavalin A and lipopolysaccharide [LPS]) and increased LPS-induced production of TNF-${\alpha}$ and IL-6, but not IL-$1{\beta}$. In conclusion, WFRG has the potential to modulate immune function and should be further investigated as an immunostimulatory agent.

Beneficial Effects of Riboflavin on Inflammatory Bowel Disease (리보플라빈의 염증성 장질환 개선 효과)

  • Sang Hee Lee;Sun Mi Hong;Mi Jeong Sung
    • Journal of the Korean Society of Food Culture
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    • v.39 no.1
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    • pp.74-81
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    • 2024
  • Ulcerative colitis (UC) is a chronic inflammatory intestinal disease characterized by an imbalance in immune function and the overexpression of inflammatory cytokines and mediators. Vitamin B2, also known as riboflavin (Libof), is an essential water-soluble vitamin with numerous beneficial properties, including antioxidant, anti-aging, anti-inflammatory, anti-nociceptive, and anti-cancer effects. In this study, we aimed to investigate the protective effects of Libof on dextran sulfate sodium (DSS)-induced experimental colitis. The C57BL/6 mice were used as the in vivo model of chronic colitis to investigate the anti-inflammatory effects of Libof. RAW 264.7 cells were used for the in vitro investigation of the molecular mechanisms underlying these effects. In vivo, Libof alleviated the DSS-induced disease activity index (DAI), colon length shortening, and colonic pathological damage. In vitro, Libof inhibited lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 production in RAW 264.7 cells. Moreover, Libof inhibited LPS-induced nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression in RAW 264.7 cells. In conclusion, these findings indicate that Libof shows potential as an agent for the treatment of UC.

Isolation and In Vitro Culture of Vascular Endothelial Cells from Mice

  • Choi, Shinkyu;Kim, Ji Aee;Kim, Kwan Chang;Suh, Suk Hyo
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.1
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    • pp.35-42
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    • 2015
  • In cardiovascular disorders, understanding of endothelial cell (EC) function is essential to elucidate the disease mechanism. Although the mouse model has many advantages for in vivo and in vitro research, efficient procedures for the isolation and propagation of primary mouse EC have been problematic. We describe a high yield process for isolation and in vitro culture of primary EC from mouse arteries (aorta, braches of superior mesenteric artery, and cerebral arteries from the circle of Willis). Mouse arteries were carefully dissected without damage under a light microscope, and small pieces of the vessels were transferred on/in a Matrigel matrix enriched with endothelial growth supplement. Primary cells that proliferated in Matrigel were propagated in advanced DMEM with fetal calf serum or platelet-derived serum, EC growth supplement, and heparin. To improve the purity of the cell culture, we applied shearing stress and anti-fibroblast antibody. EC were characterized by a monolayer cobble stone appearance, positive staining with acetylated low density lipoprotein labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate, RT-PCR using primers for von-Willebrand factor, and determination of the protein level endothelial nitric oxide synthase. Our simple, efficient method would facilitate in vitro functional investigations of EC from mouse vessels.

Antiangiogenic Activity of the Lipophilic Antimicrobial Peptides from an Endophytic Bacterial Strain Isolated from Red Pepper Leaf

  • Jung, Hye Jin;Kim, Yonghyo;Lee, Hyang Burm;Kwon, Ho Jeong
    • Molecules and Cells
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    • v.38 no.3
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    • pp.273-278
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    • 2015
  • The induction of angiogenesis is a crucial step in tumor progression, and therefore, efficient inhibition of angiogenesis is considered a powerful strategy for the treatment of cancer. In the present study, we report that the lipophilic antimicrobial peptides from EML-CAP3, a new endophytic bacterial strain isolated from red pepper leaf (Capsicum annuum L.), exhibit potent antiangiogenic activity both in vitro and in vivo. The newly obtained antimicrobial peptides effectively inhibited the proliferation of human umbilical vein endothelial cells at subtoxic doses. Furthermore, the peptides suppressed the in vitro characteristics of angiogenesis such as endothelial cell invasion and tube formation stimulated by vascular endothelial growth factor, as well as neovascularization of the chorioallantoic membrane of growing chick embryos in vivo without showing cytotoxicity. Notably, the angiostatic peptides blocked tumor cell-induced angiogenesis by suppressing the expression levels of hypoxia-inducible $factor-1{\alpha}$ and its target gene, vascular endothelial growth factor (VEGF). To our knowledge, our findings demonstrate for the first time that the antimicrobial peptides from EML-CAP3 possess antiangiogenic potential and may thus be used for the treatment of hypervascularized tumors.

Effects of Okbyungpoongsan Administration on Innate and Specific Immune Response in the Mouse (옥병풍산(玉屛風散)이 생쥐의 선천성 및 특이적 면역반응에 미치는 영향)

  • Song, Bong-Keun;Jeon, Yong-Cheol
    • The Journal of Korean Medicine
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    • v.20 no.2
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    • pp.177-186
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    • 1999
  • Okbyungpoongsan(OBPS) has long been known to have anti-allergic effect. In order to evaluate the influence on innate and specific immune response, the effects of OBPS on vascular permeability. hypersensitivities and phagocytic functions were measured. As the results, OBPS increased phagocytic activity of peritoneal macrophages in vitro and in vivo. But OBPS depressed formation of reactive oxygen intermediates(ROI) in vitro and in vivo, while the drug enhanced generation macrophages. Foot pad swelling in the mouse and contact hypersensitivity against dinitroflouorobenzene were decreased. OBPS had no effect on NK cells. But OBPS decreased vascular permeability induced by histamine without statistical significance. These results demonstrate that OBPS suppresses hypersensitivity reactions without affecting phagocytic functions and formation of ROI from macrophages. It also means that OBPS acts as a effective inducer to synthesis of nitric oxide which is effective for the infectious disease while it does damage to tissue less as it suppresses ROI, So we can conclude that OBPS could be used for the treatment of the disease related with immune function.

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in vitro Anticancer Activity and in vivo Chronic Toxicity of Homoharringtonine (개비자나무 유효성분인 homoharringtonine의 in vitro 항암활성 및 in vivo 만성 독성)

  • Yoo, Gui-Jae;Cho, Chul-Hee;Lee, Gun-Soon;Ryoo, Zae-Young;Chae, Hee-Jeong
    • Applied Biological Chemistry
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    • v.51 no.2
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    • pp.124-128
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    • 2008
  • Cell proliferation inhibitory effects of homoharringtonine (HHT), an active drug substance in Cephalotaxus koreana, against blood cancer cell line K562 were evaluated. In addition, in vivo chronic toxicity test with mouse was carried out. When K562 cell line was treated everyday for 9, 6, 3 days, $IC_{50}$ values of HHT were determined as 0.27, 0.37, and 1.10 mM respectively. The anticancer activity of HHT was comparable to adriamycin, a known anticancer drug compound for blood cancer treatment. in vivo chronic toxicity test of the HHT, the number of red blood cell (RBC) showed no significant difference. From the analysis of the liver-functional enzymes in blood, all of liver damage related enzymes such as glutamate-oxalate-transferase (GOT), glutamate-pyruvate-transferase (GPT), cholesterol (Chol) and alkaline phosphatase (ALP) showed no significant change. However, from the histologic test, a neutrophil of the band type in liver tissue was observed.

Effect of electromagnetic field exposure on the reproductive system

  • Gye, Myung-Chan;Park, Chan-Jin
    • Clinical and Experimental Reproductive Medicine
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    • v.39 no.1
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    • pp.1-9
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    • 2012
  • The safety of human exposure to an ever-increasing number and diversity of electromagnetic field (EMF) sources both at work and at home has become a public health issue. To date, many in vivo and in vitro studies have revealed that EMF exposure can alter cellular homeostasis, endocrine function, reproductive function, and fetal development in animal systems. Reproductive parameters reported to be altered by EMF exposure include male germ cell death, the estrous cycle, reproductive endocrine hormones, reproductive organ weights, sperm motility, early embryonic development, and pregnancy success. At the cellular level, an increase in free radicals and $[Ca^{2+}]i$ may mediate the effect of EMFs and lead to cell growth inhibition, protein misfolding, and DNA breaks. The effect of EMF exposure on reproductive function differs according to frequency and wave, strength (energy), and duration of exposure. In the present review, the effects of EMFs on reproductive function are summarized according to the types of EMF, wave type, strength, and duration of exposure at cellular and organism levels.

Mutational Analysis Elucidates the Role of Conserved 237 Arginine in 23S rRNA Methylation, Which is in the Concave Cleft Region of ErmSF (ErmSF에서 두 도메인 사이에 존재하는 잘 보존된 237번 아르지닌 잔기의 위치 지정 치환 변이의 효소 활성 검색을 통한 역할 규명)

  • Jin, Hyung Jong
    • Korean Journal of Microbiology
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    • v.49 no.2
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    • pp.105-111
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    • 2013
  • The Erm family of adenine-$N^6$ methyltransferases (MTases) is responsible for the development of resistance to macrolide-lincosamide-streptogramin B antibiotics through the methylation of 23S ribosomal RNA. Recently, it has been proposed that well conserved amino acids in ErnC' located in concave cleft between N-terminal 'catalytic' domain and C-terminal 'RNA-binding' domain interacts with substrate RNA. We carried out the site-directed mutagenesis and studied the function of the ErmSF R237 mutant in vitro and in vivo. R237 amino acid residue is located in the concave cleft between two domains. Furthermore this residue is very highly conserved in almost all the Erm family. Purified mutant protein exhibited only 51% enzyme activity compared to wild-type. Escherichia coli with R237A mutant protein compared to the wild-type protein expressing E. coli did not show any difference in its MIC (minimal inhibitory concentration) suggesting that even with lowered enzyme activity, mutant protein was able to efficiently methylate 23S rRNA to confer the resistance on E. coli expressing this protein. But this observation strongly suggests that R237 of ErmSF probably interacts with substrate RNA affecting enzyme activity significantly.