• 제목/요약/키워드: total cell number

검색결과 1,024건 처리시간 0.033초

Biocompatibility of Poly(MPC-co-EHMA)/Poly(L-1actide-co-glycolide) Blends

  • Gilson Khang;Park, Myoung-Kyu;Jong M. Rhee;Lee, Sang-Jin;Lee, Hai-Bang;Yasuhiko Iwasaki;Nobuo Nakabayashi;Kazuhiko Ishihara
    • Macromolecular Research
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    • 제9권2호
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    • pp.107-115
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    • 2001
  • Poly(L-lactide-co-glycolide)(PLGA) was blended with poly[$\omega$-methacryloyloxyethyl phospho-rylcholine-co-ethylhexylmethacrylate (PMEH)] (PLGA/PMEH) to endow with new functionality i.e., to improve the cell-, tissue- and blood-compatibility. The characteristics of surface properties were investigated by measurement of contact angle goniometer, Fourier-transform infrared spectroscopy with attenuated total reflectance (FTIR-ATR) and electron spectroscopy for chemical analysis (ESCA). NIH/3T3 fibroblast and bovine aortic endothelial cell were cultured on control and PLGA/PMEH surfaces for the evaluation of ceil attachment and proliferation in terms of surface functionality such as the concentration of phosphoryl-choline. Also, the behavior of platelet adhesion on PLGA/PMEH was observed in terms of the surface functionality. The contact angles on control and PLGA/PMEH surfaces decreased with increasing PMEH content from 75$^{\circ}$ to about 43$^{\circ}$. It was observed from the FTIR-ATR spectra that phosphorylcholine groups are gradually increased with increasing blended amount of MPC. The experimental P percent values from ESCA analysis were more 3.28∼7.4 times than that of the theoretical P percent for each blend films. These results clearly indicated that the MPC units were concentrated on the surface of PLGA/PMEH blend. The control and PLGA/PMEH films with 0.5 to 10.0 wt% concentration of PMEH were used to evaluate cell adhesion and growth in terms of phosphorylcholine functionality and wettability. Cell adhesion and growth on PLGA/PMEH surfaces were less active than those of control and both cell number decreased with increasing PMEH contents without the effect of surface wettability. It can be explained that the fibronectin adsorption decreased with an increase in the surface density of phosphorylcholine functional group. One can conclude the amount of the protein adsorption and the adhesion number of cells can be controlled and nonspecifically reduced by the introduction with phosphorylcholine group. Morphology of the adhered platelets on the PLGA/PMEH surface showed lower activating than control and the number of adhered platelets on the PLGA/PMEH sample decreased with increasing the phosphorylcholine contents. The amount of fibrinogen adsorbed on the PLGA/PMEH surface demonstrated that the phospholipid polar group played an important role in reducing protein adsorption on the surface. In conclusion, this surface modification technique might be effectively used PLGA film and scaffolds for controlling the adhesion and growth of cell and tissue, furthermore, blood compatibility of the PLGA was improved by blending of the MPC polymer for the application of tissue engineering fields.

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Effects of (-)-Epicatechin Gallate on porcine oocyte in vitro maturation and subsequent embryonic development after parthenogenetic activation and in vitro fertilization

  • Seo, Min-Su;So, Kyoung-Ha;Hyun, Sang-Hwan
    • 한국수정란이식학회지
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    • 제31권3호
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    • pp.153-159
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    • 2016
  • (-)-Epicatechin gallate (ECG) is a polyphenol compound of green tea exhibiting biological activities, such as antioxidant and anticancer effects. To examine the effect of ECG on porcine oocytes during in vitro maturation (IVM), oocytes were treated with 0-, 5-, 15-, and $25{\mu}M$ ECG. After maturation, we investigated nuclear maturation, intracellular glutathione (GSH) and reactive oxygen species (ROS) levels and subsequent embryonic development after parthenogenetic activation (PA) and in vitro fertilization (IVF). After 42 hours of IVM, the $5{\mu}M$ group exhibited significantly increased (p< 0.05) nuclear maturation (89.8%) compared with the control group (86.1%). However, the $25{\mu}M$ group observed significantly decreased (p< 0.05) nuclear maturation (83.5%). In intracellular maturation assessment the 5-, 15-, and $25{\mu}M$ groups had significantly increased (p< 0.05) GSH levels and decreased ROS levels compared with the controls. The 5- and $15{\mu}M$ group showed significantly increased (p< 0.05) embryo formation rates and total cell number of blastocysts after PA (18% and 68.9, 15% and 85.1 vs. 12% and 59.5, respectively) compared with controls. Although the $25{\mu}M$ group observed significantly lower blastocyst formation rates after PA (27.6% vs. 23.2%) than control group, the $5{\mu}M$ group showed significantly increased blastocyst formation rates after PA (37.2% vs. 23.2%) compared to the control group. Furthermore, the $5{\mu}M$ group measured significantly increased blastocyst formation rates (20.7% vs. 8.6%) and total cell number after IVF ($88.3{\pm}1.5$ vs. $58.0{\pm}3.6$) compared to the control group. The treatment of $5{\mu}M$ ECG during IVM affectively improved the porcine embryonic developmental competence by regulating intracellular oxidative stress during IVM.

Effect of Alpha-Linolenic Acid on Oocyte Maturation and Embryo Development in Pigs

  • Lee, Ji-Eun;Hwangbo, Yong;Kim, Hwa-Young;Lee, Won-Hee;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • 한국발생생물학회지:발생과생식
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    • 제21권2호
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    • pp.205-213
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    • 2017
  • The aim of this study was to determine the effect of additional alpha-linolenic acid (ALA) supplementation during in vitro maturation (IVM) and culture (IVC) on nucleic maturation and embryo development of pigs. Cumulus-oocyte complexes (COCs) were incubated in IVM medium containing different concentration of ALA (25, 50 and $100{\mu}M$) for 44 h. After in vitro maturation, nuclear maturation of oocytes were evaluated by aceto-orcein stain. Mature oocytes with $50{\mu}M$ ALA were fertilized and cultured in IVC medium with ALA (25, 50 and $100{\mu}M$) during early-embryogenesis (48 hours after fertilization). Then, embryos were cultured with $25{\mu}M$ ALA during early embryogenesis and/or late embryogenesis (120 hours after early-embryogenesis). In results, oocyte maturation were significantly increased by $50{\mu}M$ ALA treatment groups compared with control groups (p<0.05). Treatment of $25{\mu}M$ ALA during early-embryogenesis enhanced cleavage rate of embryo compared with other groups (p<0.05), whereas formation and total cell number of blastocyst had no significant difference. Similarly, cleavage rate of embryos were increased by $25{\mu}M$ ALA supplement during early- or late-embryogenesis than ALA treatment both stage of embryogenesis (p<0.05), but did not influence to blastocyst formation. Interestingly, total cell number of blastocyst were enhanced in ALA treatment group during early-embryogenesis. These findings indicated that ALA supplement enhance the nuclear maturation of oocyte and embryo development, however, excessive ALA could negatively influence. Therefore, we suggest that ALA is used for improvement of in vitro production of mammalian embryo and further study regarding with functional mechanism of ALA is needed.

Study of a BALB/c Mouse Model for Allergic Asthma

  • Yang, Young-Su;Yang, Mi-Jin;Cho, Kyu-Hyuk;Lee, Kyu-Hong;Kim, Yong-Bum;Kim, Jin-Sung;Kang, Myung-Gyun;Song, Chang-Woo
    • Toxicological Research
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    • 제24권4호
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    • pp.253-261
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    • 2008
  • Allergic asthma is a worldwide public health problem and a major socioeconomic burden disease. It is a chronic inflammatory disease marked by airway eosinophilia and goblet cell hyperplasia with mucus hypersecretion. Mouse models have proven as a valuable tool for studying human asthma. In the present report we describe a comparison of mouse asthma models. The experiments were designed as follows: Group I was injected with ovalbumin (OVA, i.p.) on day 1 and challenged with 1% OVA (aerosol exposure) on days $14{\sim}21$. Group II was injected on day 1, 14 and aerosol-immunized on days $14{\sim}21$. Group III was injected on day 1, 14 and immunized by 1% OVA aerosol on days $18{\sim}21$. We assessed asthma induction by determining the total number of white blood cells (WBC) and eosinophils as well as by measuring cytokine levels in bronchoalveolar lavage fluid (BALF). In addition, we evaluated the histopathological changes of the lungs and determined the concentration of immunoglobulin E (IgE) in serum. Total WBC, eosinophils, Th2 cytokines (IL-4, IL-13) and IgE were significantly increased in group I relative to the other groups. Moreover, histopathological studies show that group I mice show an increase in the infiltration of inflammatory cell-in peribronchial and perivascular areas as well as an overall increase in the number of mucus-containing goblet cells relative to other groups. These data suggest that group I can be a useful model for the study of human asthma pathobiology and the evaluation of existing and novel therapeutic agents.

Mammary Gland Indices at the End of Lactation in Javanese Thin-tail Ewes with Different Litter Sizes

  • Manalu, W.;Sumaryadi, M.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • 제11권6호
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    • pp.648-654
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    • 1998
  • Twenty-four Javanese thin-tail ewes (11, 9, and 4 ewes giving birth to 1, 2, and 3 lambs, respectively) with similar body weight and age at breeding were used to study serum progesterone concentrations during pregnancy, milk production during lactation, and mammary gland indices at the end of lactation (3 months postpartum). The results of the experiment showed that averages serum progesterone concentrations during pregnancy in the ewes giving birth to twin and triplet lambs were higher (p < 0.01) than those giving birth to a single lamb. Ewes giving birth to 3 lambs had higher (p < 0.01) mammary dry fat-free tissue (DFFT) (by 31 and 34%), DNA concentration (by 25 and 16%) and RNA concentration (by 29 and 16%) at the end of lactation than those giving birth to 1 and 2 lambs. There was no difference in mammary collagen, protein and glycogen concentrations at the end of lactation among litter sizes. Ewes giving birth to 3 lambs had higher (p < 0.01) total mammary DNA content (by 64 and 61%) and RNA content (by 69 and 53%) at the end of lactation than those giving birth to 1 and 2 lambs. There was no difference in total mammary collagen, protein and glycogen contents at the end of lactation among litter sizes. Even though ewes with higher litter size had numerically higher milk production, there was no significant difference in milk production per 4 h among litter sizes. The results of the experiment indicated that ewes having higher litter size had greater mammary cell number and synthetic activities at the end of lactation. The results suggested that ewes with higher progesterone concentrations and better developed mammary glands during pregnancy could maintain higher cell number and activities throughout lactation.

Effects of High Dose Lysophosphatidic Acid Supplement during IVC on Preimplantation Development of Porcine Embryos

  • Jin, Minghui;Yu, Il-Jeoung;Jeon, Yubyeol
    • 한국수정란이식학회지
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    • 제32권4호
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    • pp.275-285
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    • 2017
  • Lysophosphatidic acid (LPA) is an important signaling molecule. Here, the effect and mechanism of LPA on the preimplantation development of porcine embryos during in vitro culture (IVC) was examined. Porcine embryos were cultured in porcine zygote medium (PZM-3) supplemented with $30{\mu}M$ LPA during different days. There was a significantly higher cleavage rate in Day 1-7 and significantly higher total cell number of blastocysts in Day 1-3 and Day 4-7. It was also found that messenger RNA (mRNA) expression level of PCNA, BCL2 and BAX in blastocysts obtained from D1-7 group were significantly higher and BCL2/BAX mRNA ratio in D1-3 group was significantly lower than control group but Day 4-7 and Day 1-7 groups were comparable with control group. Treatment with $20{\mu}M$ PLC inhibitor significantly decreased the embryo cleavage rate and blastocyst formation rate. Moreover, LPA as an activator of PLCs, enhanced the $30{\mu}M$ LPA + $20{\mu}M$ U73122 group embryo cleavage rate which similar with control group. In conclusion, the results suggest that treatment with LPA during IVC improves the porcine early embryo cleavage by activation of PLC signaling pathway and regulate the mRNA expression that contribute to total cell number of blastocysts during blastocyst formation.

중금속 오염이 양서류 시원생식세포 발생에 미치는 영향 (Effects of the Heavy Metal Pollution on the Primordial Germ Cells of Developing Amphibia)

  • Hah, Jae-Chung
    • 한국동물학회지
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    • 제21권2호
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    • pp.43-58
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    • 1978
  • 양서류 수정란을 제 1 난할전에 중금속으로 처리하였던바 개구리의 시원생식세포수의 양전변화를 가져 왔다. 중금속의 일정량의 처리는 양서류 초기발생중에 시원생식세포 형성을 저하시켰다. 납 70ppm과 카드뮴 4ppm 이상에서 배양하면 9내지 12mm배에서 시원생식세포의 수적감소를 가져 왔으나 그후의 발생에서는 거의 정상군과같은 증가율을 보였다. 한편 수은 0.8ppm이상의 처리군에서는 생식 세포형성에 보다 심한 저하를 가져 왔으며 그후의 증가율도 대체로 낮았다. 위의 사실들은 자외선 조사 실험에서 얻은 결과와는 달리 양서류란에 있어서 중금속 처리는 조직에 심한 손상을 주는 다량투여에도 시원생식세포의 완전 배제에는 효과가 적다는 것을 암시한다.

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김치의 급여가 흰쥐의 체내 지질함량과 비장 면역세포 증식능력에 미치는 영향 (The Effects of Kimchi Intake on Lipid Contents of Body and Mitogen Response of Spleen Lymphocytes in Rats)

  • 김지연;이연숙
    • 한국식품영양과학회지
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    • 제26권6호
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    • pp.1200-1207
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    • 1997
  • Effects of kimchi on lipid metabolism and immune function were studied in experiments using 63mals SD rats fed 6 inds of Baechu-kimchi containing diet during 4 weeks. Three kinds of freeze dried kimchi differ in fermentation period (not fermented, 3-, 6-week-fermented at 4$^{\circ}C$) were added at 5%, 10% of the diet containing 15% lard. The levels of serum total lipid and triglyceride and the content of liver total lipid and triglyceride of all kimchi groups were lower than those of a control group. But the levels of serum and liver cholesterol is not affected by kimchi intake. The triglyceride concentration of epididymal fat pad and feces of kimchi groups were higher than those of a control group. The food efficiency ratio, epididymal fat pad weight of 3-, 6-week-fermented kimchi 10% groups were significantly lower than control and not-fermented kimchi groups. Especially 6-week-fermented kimchi groups showed adipocytes, less in number and larger in size than those of other groups. The blastogenesis of spleen lymphocytes to LPS was higher in rats fed fermented kimchi diets than rats fed control and not-fermented kimchi diet. These results suggest that kimchi stimulates lipid mobilization to epididymal fat pad and lipid excretion via feces, so lower serum and liver triglyceride concentration. The fermented kimchi stimulate the proliferation of B cell and lower the lipid accumulation in epididymal fat pad, especially kimchi fermented for 6 weeks at 4$^{\circ}C$ lower the adipose cell number.

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Protodioscin protects porcine oocytes against H2O2-induced oxidative stress during in vitro maturation

  • So-Hee Kim;Seung-Eun Lee;Jae-Wook Yoon;Hyo-Jin Park;Seung-Hwan Oh;Do-Geon Lee;Da-Bin Pyeon;Eun-Young Kim;Se-Pill Park
    • Animal Bioscience
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    • 제36권5호
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    • pp.710-719
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    • 2023
  • Objective: The present study investigated whether protodioscin (PD), a steroidal saponin mainly found in rhizome of Dioscorea species, alleviates oxidative stress-induced damage of porcine oocytes during in vitro maturation. Methods: Oocytes were treated with different concentrations of PD (0, 1, 10, 100, and 200 µM) in the presence of 200 µM H2O2 during in vitro maturation. Following maturation, spindle morphology and mitogen-activated protein kinase activity was assessed along with reactive oxygen species level, GSH activity, and mRNA expression of endogenous antioxidant genes at the MII stage. On the day 7 after parthenogenetic activation, blastocyst formation rate was calculated and the quality of embryo and mRNA expression of development-related genes was evaluated. Results: Developmental competence was significantly poorer in the 0 µM PD-treated (control) group than in the non-treated (normal) and 10 µM PD-treated (10PD) groups. Although the reactive oxygen species level did not significantly differ between these three groups, the glutathione level and mRNA expression of antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, nuclear factor erythroid 2-related factor 2 [Nrf2], and hemo oxygenase-1 [HO-1]) were significantly higher in the normal and 10PD groups than in the control group. In addition, the percentage of oocytes with defective spindle and abnormal chromosomal alignment was significantly lower and the ratio of phosphorylated p44/42 to total p44/42 was significantly higher in the normal and 10PD groups than in the control group. The total cell number per blastocyst was significantly higher in the 10PD group than in the control group. The percentage of apoptotic cells in blastocysts was highest in the control group; however, the difference was not significant. mRNA expression of development-related genes (POU domain, class 5, transcription factor 1 [POU5F1], caudal type homeobox 2 [CDX2], Nanog homeobox [NANOG]) was consistently increased by addition of PD. Conclusion: The PD effectively improves the developmental competence and quality of blastocysts by protecting porcine oocytes against oxidative stress.

신선초 에탄올 추출물이 인체 유방암 MDA-MB-231 세포에서 세포증식, 세포사멸과 ROS 축적에 미치는 영향 (The Effect of Angelica keiskei Ethnol Extract on Proliferation, Apotosis and ROS Accumulation in Human Breast Cancer MDA-MB-231 Cells)

  • 정유진;남미경;강금지
    • 동아시아식생활학회지
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    • 제21권1호
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    • pp.24-30
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    • 2011
  • The anti-cancer effects of Angelica keiskei ethanol extract were evaluated in human breast cancer MDA-MB-231 cells. The concentrations of extract were 1, 2, 3, 4 and 5 mg/mL. Dose-dependent reductions in the number of cells with altered cell shape and pyknotic nuclei were observed at 48 h after treatments. MTT assay also exhibited a similar dose-dependent reduction in mitochondrial reductase activity (p<0.05), in particular, with a rapid reduction in the activity of the 5 mg/mL group. Analysis of cell death with propidium iodide (PI) staining revealed only a slight increase in cell death in the 5 mg/mL group. Analysis of bromodeoxyuridine (BrdU) incorporations also showed a dose-dependent reduction in cell proliferation (p<0.05). Finally, increases in total radical oxygen species (ROS) accumulation in cells, as revealed by DCF-DA staining, were observed in the treated groups in a similar dose-dependent fashion (p<0.05). These results indicate that Angelica keiskei ethanol extract exhibiting anti-cancer effects in MDA-MB-231 cells causes multiple changes in cell shape, enzyme activity, and ROS accumulation, thereby inducing cell death.