• Title/Summary/Keyword: 3D co-culture

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Mouse Strain-Dependent Osteoclastogenesis in Response to Lipopolysaccharide

  • Choi, Ho-Gil;Kim, Jin-Moon;Kim, Bong-Ju;Yoo, Yun-Jung;Cha, Jeong-Heon
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.566-571
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    • 2007
  • Bacterial lipopolysaccharide (LPS) is a potent stimulator of bone resorption in periodontitis. Co-culture systems of mouse calvaria-derived osteoblasts and bone marrow-derived preosteoclasts were used as an in vitro osteoclast differentiation. This study revealed that co-cultures using ddY or ICR mouse strain responded differently to LPS while responded equally to $1{\alpha},25(OH)_2D_3$. Thus, the different response to LPS indicates dissimilarity of two mouse stains in their capacity for generating osteoclasts while the two mouse strains share the similarity in response to $1{\alpha},25(OH)_2D_3$. To identify which cells between osteoblasts and preosteoclasts in the co-culture are responsible for the dissimilarity, the reciprocal co-cultures were performed between ddY and ICR mouse strains. The treatment of $1,25(OH)_2D_3$ to ddY/ICR (osteoblasts from ddY/preosteoclasts from ICR) and ICR/ddY reciprocal co-cultures also showed the similarity. In case of LPS treatment, the results of ddY/ICR were similar to ddY/ddY and the results of the other reciprocal co-culture, ICR/ddY combination, were consistent with those of ICR/ICR. It suggests that the dissimilarity between the two mouse strains may resident in osteoblasts but not in preosteoclasts. Therefore, the osteoblast is responsible for mouse strain-dependent osteoclastogenesis in response to LPS. Although mouse models will continue to provide insights into molecular mechanisms of osteoclastogenesis, caution should be exercised when using different mouse strains, especially ddY and ICR strains as models for osteoclast differentiation.

Anti-Cancer Effects of Peptides Purified from Culture Supernatant of Lactobacillus casei (Lactobacillus casei의 배양물에서 분리한 물질의 항암 효과)

  • Kim, Jung-Hwa;Kim, Dong-Myung;Baek, Hong;Lee, Seung-Hoon;Chung, Myung-June
    • Journal of Dairy Science and Biotechnology
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    • v.26 no.1
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    • pp.5-10
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    • 2008
  • This study was conducted to isolate protein components from culture supernatant of Lactobacillus casei. and measure anti-cancer activity. The protein components were isolated A and B on Ultrafiltration membrane(3, 10, 30, 100 KDa). And the protein components A and B were isolated fractions(number $3{\sim}9$) on FPLC. Experimental studies were progressed through the cell cytotoxicity and anti-cancer activities. Cell cytotoxicity test using human kidney normal cell(293) showed cytotoxicity of below 20% by the protein components A and B($100{\mu}g/mL$). The anti-cancer activity was increased up to 70% by the protein components A and B($100{\mu}g/mL$) in AGS(stomach cancer), A549(lung cancer), MCF-7 (breast cancer), SK-OV-3(ovary cancer) and LoVo(colon cancer). Cell cytotoxicity test was showed cytotoxicity of about 50% by the fractions(number 3, 8, 9) isolated FPLC. The others have not the cytotoxicity about the human normal cell. The anti-cancer activity was increased up to 70% by the fraction number 7 in cancer cell line. Therefore the components isolated from culture supernatant of Lactobacillus casei were showed anti-cancer activity.

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Development of Intelligent Traditional Culture Retrieval System based on 3D Digital Timeline (3D 디지털 연표 기반의 지능형 전통문화 검색 시스템 개발)

  • Shin, Yutak;Jo, Jaechoon
    • Journal of Convergence for Information Technology
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    • v.9 no.10
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    • pp.154-162
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    • 2019
  • Despite the development of information and communication technology, which has a great impact on society and culture, there is no platform that provides a systematic classification and state-of-the-art information retrieval system on the traditional culture. Therefore, this paper developed a traditional culture retrieval system capable of convergence services with systematic classification and retrieval based on automatically generate and visualize the 3D timeline. This system provides the function of collecting traditional culture contents, classifying and storing collected traditional culture contents, and automatically generating 3D digital timeline based on stored traditional culture contents. In addition, a system satisfaction questionnaire was developed to evaluate the usability of the system, and 19 students participated in verifying the system. As a result of the experiment, the satisfaction of the system showed that all items were 'satisfied' on average.

Early Mouse Embryonic Development In Vitro by Co-culture with Bovine and Porcine Oviductal Epithelial Cells (소와 돼지의 난관 상피세포와의 공배양이 마우스 초기배의 체외발달에 미치는 영향)

  • Lee, S.;Hur, E. J.;Seok, H. B.
    • Journal of Embryo Transfer
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    • v.11 no.3
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    • pp.201-210
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    • 1996
  • This experiment was carried out to evaluate the effect of early mouse embryonic development in vitro by co-culture with bovine and porcine oviductal epithelial cells (BOEC and POEC). The 2-cell embryos were collected from the oviducts of the superovulated and mated cultured in D-PBS /15% FCS at 48 hours after hCG injection. The in vitro developmental rate of blastocyst formation in the embryos were examined under the fllowing treatments; 1) TCM 199 added 15% HCS, 2) Ham's F-10 added 15% HCS, 3) MediCult IVF medium, 4) TCM 199 added 15% HCS + BOEC, 5) TCM 199 added 15% HCS + POEC, 6) Ham's F40 added 15% HCS + BOEC, 7) Ham's F-10 added 15% HCS + POEC,8) MediCult IVF medium + BOEC, 9) MediCult IVF medium + POEC. For a comparative study of in vitro development for 96 hours after hCG injection, were cultured with oviductal epithelial cell and media only. The obtained results were 2-cell embryos developed to the blastocyst stage in TCM 199, Ham's F-10 and MediCult IVF medium at the rates of 84.4,83.2 and 81.6%. respectively. The higher developmental rates(91~97%) of blastocyst formation was appeared when the embryos were co-cultured with a monolayer of bovine or porcine oviductal epithelial cells in TCM 199 or Ham's F-10 and MediCult IVF media. No significant difference in developmental rates was shown between bovine and porcine oviductal epithelial cells but significant difference in co-culture system in comparison between media only system and co-cultures. In conclusions, oviductal epithelial cells, BOEC and POEC, when co-culture with mouse early embryos improved the rates of development, blastocyst and hatching. Therefore, it is suggested that co-culture system using oviductal epithelial cells improve early embryonic developtnent in mouse.

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Hypertonicity Down-regulates the $1{\alpha},25(OH)_2$ Vitamin $D_3$-induced Osteoclastogenesis Via the Modulation of RANKL Expression in Osteoblast

  • Jeong, Hyun-Joo;Yushun, Tian;Kim, Bo-Hye;Nam, Mi-Young;Lee, Hyun-A;Yoo, Yun-Jung;Seo, Jeong-Taeg;Shin, Dong-Min;Ohk, Seung-Ho;Lee, Syng-Ill
    • International Journal of Oral Biology
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    • v.30 no.1
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    • pp.23-30
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    • 2005
  • Bone remodeling is a process controlled by the action of two major bone cells; the bone forming osteoblast and the bone resorbing osteoclast. In the process of osteoclastogenesis, stromal cells and osteoblast produce RANKL, OPG, and M-CSF, which in turn regulate the osteoclastogenesis. During the bone resorption by activated osteoclasts, extracellular $Ca^{2+}/{PO_4}^{2-}$ concentration and degraded organic materials goes up, providing the hypertonic microenvironment. In this study, we tested the effects of hypertonicity due to the degraded organic materials on osteoclastogenesis in co-culture system. It was examined the cellular response of osteoblastic cell in terms of osteoclastogenesis by applying the sucrose, and mannitol, as a substitute of degraded organic materials to co-culture system. Apart from the sucrose, mannitol, and NaCl was tested to be compared to the effect of organic osmotic particles. The addition of sucrose and mannitol (25, 50, 100, 150, or 200 mM) to co-culture medium inhibited the number of tartrate-resistant acid phosphatase (TRAP) positive multinucleated cells induced by 10 nM $1{\alpha},25(OH)_2vitaminD_3$ ($1{\alpha},25(OH)_2D_3$). However, NaCl did exert harmful effect upon the cells in this co-culture system, which is attributed to DNA damage in high concentration of NaCl. To further investigate the mechanism by which hypertonicity inhibits $1{\alpha},25(OH)_2D_3$-induced osteoclastogenesis, the mRNA expressions of receptor activator of nuclear factor (NF)-kB ligand (RANKL) and osteoprotegerin (OPG) were monitored by RT-PCR. In the presence of sucrose (50 mM), RANKL mRNA expression was decreased in a dose-dependent manner, while the change in OPG and M-CSF mRNA were not occurred in significantly. The RANKL mRNA expression was inhibited for 48 hours in the presence of sucrose (50 mM), but such a decrement recovered after 72 hours. However, there were no considerable changes in the expression of OPG and M-CSF mRNA. Conclusively, these findings strongly suggest that hypertonic stress down-regulates $1{\alpha},25(OH)_2D_3$-induced osteoclastogenesis via RANKL signal pathway in osteoblastic cell, and may playa pivotal role as a regulator that modulates osteoclastogenesis.

Effects of Trophoblastic Vesicle and Estradiol-$17\beta$ on the Development in Vitro of Rabbit Embryos (Trophoblastic Vesicle과 Estradiol-$17\beta$의 첨가가 가토배의 발달에 미치는 영향)

  • 오하식;박충생
    • Korean Journal of Animal Reproduction
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    • v.10 no.1
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    • pp.76-82
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    • 1986
  • This experiment was conducted to determine the effects of trophoblastic vesicles (TV) and estradiol-17$\beta$ on the development in vitro of rabbit embryos. Thirty matured female rabbits were treated with PMSG followed by HCG injection and mating. Embryos were recovered with D-PBS (Dulbecco's Phosphate Buffered Saline) after superovulation, and normally developed to two-to four-cell embryos were used in the subsequent in vitro culture. Basal medium was Medium-199 su, pp.emented with 1.5% bovine serum albumin. Embryo on Day 5 after mating (Day 0) was cut into two or three pieces to remove the embryonic disc. Each piece of tissue was cultured for 24 hours at 37$^{\circ}C$ in 0.5 mlMedium-199 in 5% CO2. During culture, peices of trophoblastic tissue changed into spherical vesicles which were used for co-culture. These spheres were called trophoblstic vesicles. Two-to four-cell embryos were cultured for 4 days in Medium-199 in the absence or presence of trophoblastic vesicle, and two-to four-cell embryos cultured with varing concentration (0, 0.1, 1, 10ng/ml) of estradiol-17$\beta$ for 4 dyas. Culture vessels used were watch glass for coculture with trophoblastic vesicles and micortube for estradiol-17$\beta$ infusion. Compared with the Medium-199 alone as basal culture medium, more blastocysts (46.7% vs 15.1%; P<0.01) and morulae (84.4% vs 56.6%; P<0.05) were developed in the co-culture with trophoblastic vesicles. Estradiol-17$\beta$ infused in culture medium was not effective for embryo development to blastocysts (78.3% in control, 50.0% in 0.1ng/ml, 61.5% in 1ng/ml and 64.4% in 10ng/ml) and also to morulae (91.3% in control, 84.2% in 0.1ng/ml, 92.3% in 1ng/ml and 91.1% in 10ng/ml). Compared with the watch glass culture mehotd, more (P<0.01) blastocysts were developed in microtube culture (78.3% vs 56.6%) and more (P<0.01) morulae in microtube culture (91.3% vs 56.6%).

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Antitumor and immunomodulatory effects of a hot water extract of Phellinus baumii using Sarcoma-180 in the mouse (Pheflinus baumii 자실체 추출물의 Sarcoma-180 에 대한 항암 및 면역효과)

  • Ha, Hyo-Cheol;Kim, Hyun-Pyo;Sim, Ji Young;Chang, Yun-Hee;Kim, Hyun Su
    • Journal of Mushroom
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    • v.2 no.4
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    • pp.169-174
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    • 2004
  • A hot water extract(HWE-P4) was separated from the fruit bodies of PMO-P4, and its antitumor and immunomodulatory activities against sarcoma-180 in ICR mice were investigated. The internal transcribed spacer(ITS) regions from PMO-P4 were amplified using polymerase chain reaction(PCR) and sequenced. The results revealed that PMO-P4 was belong to the Phellinus baumii. When oral administration at the dose of 160mg/kg/day in the mice until the end of the experiment with 2 week's pre-feeding of the HWE-P4, the survival rate of the mice was 152% for 50days after the inoculation of sarcoma-180 and the suppression rate of the tumor growth was 35.3%(p<0.05) for 28 days after inoculation of sarcoma-180. The HWE-P4 increased 71.4% of the CD4/CD8 ratio and 5-fold of the expression of CD25(IL-2 receptor chain) compared with the control. From these results, the antitumor activity of HWE-P4 is exerted through its immunomodulating activity on the host's immune system.

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Commercial Production of Seed Garlic by Tissue Culture Technique (조직배양에 의한 씨마늘의 상업적 생산)

  • Nam, Sang-Il;Park, Ju-Hyun;Choi, Jong-In;Kwon, Ki-Seok;Uhm, Jeong-Sik
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.171-177
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    • 2002
  • We, Tong Yang Moolsan Co. Ltd. (TYM) set up the mass-production system for virus-free seed garlic via tissue culture technique. TYM's tissue culture technique is called as 'Multiple shoot propagation technique' This technique can lead mass propagation of genetically homogeneous seed garlic in a short period because of its highly proliferation rate of in vitro shoots (15/sup 10//year). TYM researchers applied the technique to some selected garlic cultivars with superior characteristics and carried out field test of productivity in the inside and outside of the country for several years. According to the yearly results of field test with virus-free seed garlic, we ascertained that virus-free seed garlic can produce the highly yield increase (max. above 50%) and also can enhance the product quality. Consequently, we estimated that TYM's seed garlic will contribute to farmers with increase of income and can elevate the national position of garlic market in the world for its competitive power of technical and production cost.

Keratinocyte Migration in a Three-Dimensional In Vitro Wound Healing Model Co-Cultured with Fibroblasts

  • Iyer, Kritika;Chen, Zhuo;Ganapa, Teja;Wu, Benjamin M.;Tawil, Bill;Linsley, Chase S.
    • Tissue Engineering and Regenerative Medicine
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    • v.15 no.6
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    • pp.721-733
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    • 2018
  • BACKGROUND: Because three-dimensional (3D) models more closely mimic native tissues, one of the goals of 3D in vitro tissue models is to aid in the development and toxicity screening of new drug therapies. In this study, a 3D skin wound healing model comprising of a collagen type I construct with fibrin-filled defects was developed. METHODS: Optical imaging was used to measure keratinocyte migration in the presence of fibroblasts over 7 days onto the fibrin-filled defects. Additionally, cell viability and growth of fibroblasts and keratinocytes was measured using the $alamarBlue^{(R)}$ assay and changes in the mechanical stiffness of the 3D construct was monitored using compressive indentation testing. RESULTS: Keratinocyte migration rate was significantly increased in the presence of fibroblasts with the cells reaching the center of the defect as early as day 3 in the co-culture constructs compared to day 7 for the control keratinocyte monoculture constructs. Additionally, constructs with the greatest rate of keratinocyte migration had reduced cell growth. When fibroblasts were cultured alone in the wound healing construct, there was a 1.3 to 3.4-fold increase in cell growth and a 1.2 to 1.4-fold increase in cell growth for keratinocyte monocultures. However, co-culture constructs exhibited no significant growth over 7 days. Finally, mechanical testing showed that fibroblasts and keratinocytes had varying effects on matrix stiffness with fibroblasts degrading the constructs while keratinocytes increased the construct's stiffness. CONCLUSION: This 3D in vitro wound healing model is a step towards developing a mimetic construct that recapitulates the complex microenvironment of healing wounds and could aid in the early studies of novel therapeutics that promote migration and proliferation of epithelial cells.