• Title/Summary/Keyword: in vivo maturation

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Differential expression of αB-crystallin causes maturation-dependent susceptibility of oligodendrocytes to oxidative stress

  • Kim, Ji Young;Lee, Eun Young;Sohn, Hyun Joon;Kim, Si Wook;Kim, Chan Hyung;Ahn, Hee Yul;Kim, Dong Woon;Cho, Sa Sun;Seo, Je Hoon
    • BMB Reports
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    • v.46 no.10
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    • pp.501-506
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    • 2013
  • Oligodendrocyte precursor cells (OPCs) are most susceptible to oxidative stress in the brain. However, the cause of differences in susceptibility to oxidative stress between OPCs and mature oligodendrocytes (mOLs) remains unclear. Recently, we identified in vivo that ${\alpha}B$-crystallin (aBC) is expressed in mOLs but not in OPCs. Therefore, we examined in the present study whether aBC expression could affect cell survival under oxidative stress induced by hydrogen peroxide using primary cultures of OPCs and mOLs from neonatal rat brains. Expression of aBC was greater in mOLs than in OPCs, and the survival rate of mOLs was significantly higher than that of OPCs under oxidative stress. Suppression of aBC by siRNA transfection resulted in a decrease in the survival rate of mOLs under oxidative stress. These data suggest that higher susceptibility of OPCs than mOLs to oxidative stress is due, at least in part, to low levels of aBC expression.

Oocyte maturity in repeated ovarian stimulation

  • Lee, Jae-Eun;Kim, Sang-Don;Jee, Byung-Chul;Suh, Chang-Suk;Kim, Seok-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.4
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    • pp.234-237
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    • 2011
  • Objective: During stimulated IVF cycles, up to 15% of oocytes are recovered as immature. The purpose of this study was to investigate the trend of oocyte maturity in repeated ovarian stimulation for IVF. Methods: One hundred forty-eight patients were selected who underwent two consecutive IVF cycles using same stimulation protocol during 2008 to 2010. Ovarian stimulation was performed with FSH and human menopausal gonadotropin and flexible GnRH antagonist protocol in both cycles. Oocyte maturity was assessed according to presence of germinal vesicle (GV) and the first polar body. Immature oocyte was defined as GV stage or metaphase I oocyte (GV breakdown with no visible polar body) and cultured up to 48 hours. If matured, they were fertilized with ICSI. Results: Percentages of immature oocytes were 30.8% and 32.9% ($p$=0.466) and IVM rates of immature oocytes were 36.2% and 25.7% ($p$=0.077), respectively. A significant correlation was noted between percentage of immature oocytes in the two cycles (R=0.178, $p$=0.03). Women with >40% immaturity in both cycles (n=21) showed lower fertilization rate of $in$ $vivo$ matured oocytes (56.4% vs. 72.0%, $p$=0.005) and lower pregnancy rate (19.0% vs. 27.1%, $p$=0.454) after the second cycle when compared with women with <40% immaturity (n=70). In both groups, female age, number of total retrieved oocyte and embryos transferred were similar. Conclusion: In repeated ovarian stimulation cycles for IVF, the immature oocyte tended to be retrieved repetitively in consecutive IVF cycles.

Effect of Fibroblast Growth Factor 23 on Osteoblastic Differentiation and Mineralization of D1 Mesenchymal Stem Cells (섬유모세포성장인자-23이 D1 간엽줄기세포에서 조골세포로의 분화 및 기질 광화에 미치는 영향)

  • Park, Kyeong-Lok
    • Journal of Life Science
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    • v.26 no.3
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    • pp.331-337
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    • 2016
  • Although fibroblast growth factor 23 (FGF23) is exclusively produced in osteoblasts and osteocytes, its main target is the kidney, where it decreases phosphate reabsorption by suppressing Na-phosphate cotransporters. Independently of its action on phosphate homeostasis, FGF23 also inhibits bone formation in vivo. In a calvarial osteoblastic cell model, FGF23 was shown to negatively affect extracellular matrix mineralization. This study investigated whether FGF23 had similar effects on osteoblast maturation, including differentiation and mineralization of bone marrow-derived mesenchymal stem cells (MSCs). D1 MSCs were cultured in an osteogenic medium containing β-glycerophosphate, ascorbic acid, and dexamethazone. Osteoblastic differentiation was evaluated by alkaline phosphatase (Alp) staining, and matrix mineralization was evaluated by alizarin red staining and calcium deposition. The expression of differentiation-stimulating genes Runx2, Alp, and osteocalcin and mineralization-inhibiting genes Enpp1 and Ank was analyzed using semiquantitative RT-PCR. Supraphysiological doses of FGF23 did not stimulate proliferation or osteoblastic differentiation of MSCs. Matrix mineralization 1, 2, and 3 weeks after the FGF23 treatment did not vary between control and FGF23 groups, although time-dependent enhancement of mineralization was obvious. Calcium deposition was also unchanged after the FGF23 treatment. mRNA expression levels of differentiation- and mineralization-related genes were also similar between the groups. Despite these negative findings, FGF23 signaling through FGF receptors seemed to function normally, with phosphorylation of the Erk protein more evident in the FGF23 group than in controls. These findings suggest that unlike calvarial osteoblasts, FGF23 is not likely to affect osteoblastic differentiation and mineralization of MSCs.

Comparative analysis of the in vivo kinetic properties of various bone substitutes filled into a peri-implant canine defect model

  • Jingyang Kang;Masaki Shibasaki;Masahiko Terauchi;Narumi Oshibe;Katsuya Hyodo;Eriko Marukawa
    • Journal of Periodontal and Implant Science
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    • v.54 no.2
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    • pp.96-107
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    • 2024
  • Purpose: Deproteinized bovine bone or synthetic hydroxyapatite are 2 prevalent bone grafting materials used in the clinical treatment of peri-implant bone defects. However, the differences in bone formation among these materials remain unclear. This study evaluated osteogenesis kinetics in peri-implant defects using 2 types of deproteinized bovine bone (Bio-Oss® and Bio-Oss/Collagen®) and 2 types of synthetic hydroxyapatite (Apaceram-AX® and Refit®). We considered factors including newly generated bone volume; bone, osteoid, and material occupancy; and bone-to-implant contact. Methods: A beagle model with a mandibular defect was created by extracting the bilateral mandibular third and fourth premolars. Simultaneously, an implant was inserted into the defect, and the space between the implant and the surrounding bone walls was filled with Bio-Oss, Bio-Oss/Collagen, Apaceram-AX, Refit, or autologous bone. Micro-computed tomography and histological analyses were conducted at 3 and 6 months postoperatively (Refit and autologous bone were not included at the 6-month time point due to their rapid absorption). Results: All materials demonstrated excellent biocompatibility and osteoconductivity. At 3 months, Bio-Oss and Apaceram-AX exhibited significantly greater volumes of formation than the other materials, with Bio-Oss having a marginally higher amount. However, this outcome was reversed at 6 months, with no significant difference between the 2 materials at either time point. Apaceram-AX displayed notably slower bioresorption and the largest quantity of residual material at both time points. In contrast, Refit had significantly greater bioresorption, with complete resorption and rapid maturation involving cortical bone formation at the crest at 3 months, Refit demonstrated the highest mineralized tissue and osteoid occupancy after 3 months, albeit without statistical significance. Conclusions: Overall, the materials demonstrated varying post-implantation behaviors in vivo. Thus, in a clinical setting, both the properties of these materials and the specific conditions of the defects needing reinforcement should be considered to identify the most suitable material.

Entinostat, a histone deacetylase inhibitor, increases the population of IL-10+ regulatory B cells to suppress contact hypersensitivity

  • Min, Keun Young;Lee, Min Bum;Hong, Seong Hwi;Lee, Dajeong;Jo, Min Geun;Lee, Ji Eon;Choi, Min Yeong;You, Jueng Soo;Kim, Young Mi;Park, Yeong Min;Kim, Hyuk Soon;Choi, Wahn Soo
    • BMB Reports
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    • v.54 no.10
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    • pp.534-539
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    • 2021
  • IL-10+ regulatory B (Breg) cells play a vital role in regulating the immune responses in experimental autoimmune encephalomyelitis, colitis, and contact hypersensitivity (CHS). Several stimulants such as lipopolysaccharide (LPS), CD40 ligand, and IL-21 spur the activation and maturation of IL-10+ Breg cells, while the epigenetic mechanism for the IL-10 expression remains largely unknown. It is well accepted that the histone acetylation/deacetylation is an important mechanism that regulates the expression of IL-10. We found that entinostat, an HDAC inhibitor, stimulated the induction of IL-10+ Breg cells by LPS in vitro and the formation of IL-10+ Breg cells to suppress CHS in vivo. We further demonstrated that entinostat inhibited HDAC1 from binding to the proximal region of the IL-10 expression promoter in splenic B cells, followed by an increase in the binding of NF-κB p65, eventually enhancing the expression of IL-10 in Breg cells.

Studies on the Early Development of the Mouse Embryo Transplanted in the Anterior Chamber of the Eye (眼前房내에 이식한 생쥐배의 초기발생에 관한 연구)

  • Cho, Wan Kyoo;Kim, Moon Kyoo
    • The Korean Journal of Zoology
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    • v.15 no.1
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    • pp.25-33
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    • 1972
  • Two-Cell mouse embryos were incubated in the anterior chamber of the rat eye, which has been known as the best place among other animals' for the mouse ovum maturation, in order to observe the capability of their early development. Within 120 hours after incubation, 71.0% of two-cell embryos have developed to the blastocysts in the male rat eye, while only 38.5% in the eye of the same mouse as donated two-cell embryos. Thus, the rat eye chamber provides more favourable environment to the embryos than the mouse itself. The results are consistent with those of the previous studies comparing the maturation of the mouse follicular oocytes in the mouse and the rat eye chamber. Although the aqueous humor which is filled in the anterior chamber of the eye is characterized by its specific properties, being of higher osmolarity, higher concentrations of ascorbic acid, pyruvate and lactate, but lower of proteins and lower temperature than those in blood or lymph serum, The embryos are able to under-take their cleavage as normal as in vivo or in vitro. Concerning with a number of studies in vitro on the development of the mouse embryos which are requiring a very limited condition, the fact that they are able to manage their further development under very different enviroment from our knowledges would provide us a moment to understand their behavior during the early development. The difference of the proportion of the developed blastocysts between in the mouse eye chamber and in the rat can possibly be resulted from the species specific difference in the physicochemical properties between their eye chambers. This assumption is based upon the findings by many investigators who chmpared the nature of the eye chamber of various animals. As a consequence, the rat eye chamber might consist of better properties for the embryonal growth than the mouse eye chamber. The mouse embryos cleaved with a delayed period. In normal development they complete almost the cleavage within 94 hours after fertilization. However, in the present studies, 81.1% of two-cell embryos developed to the blastocysts and the morula in 120 hours in the eye chamber, assumed to be about 154 hours after fertilization. Such delay in development would be caused mainly by the low temperature of the eye chamber. At present we can make two assumptions to explain the capability of the emtryonal development in the eye chambers. One is that the embryos would possess an ability to adapt themselves to the environment which provides unfavourable conditions. The other is that the embryos might remain for a certain duration in the eye chamber, which is filled with a new body fluid produced immediately after the loss of the aqueous humor and the fluid of which becomes similar to blood serum in component. The first assumption is highly reliable since the embryonal cells are mostly at the undifferentiated state and so they probably engage a simple metabolism during their early period. The second assumption is induced by the fact that the rabbit eye chamber produces a plasmoid humor which has mostly similar components to blood serum after loss of aqueous humor through cornea by puncturing. However, the plasmoid humor is substituted by the initial aqueous humor in eight hours. Even though this finding, production of the new fluid, could be applied to the rat eye, it is hardly reliabel that the plasmoid humor remains for such a long period as 120 hours. Consequently, the development of the embryos is more likely due to their adaptability to the new environment during their early developmental stages.

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Developmental Characteristics of SCNT Pig Embryos Knocked-out of Alpha-1,3-Galactosyltransferase Gene

  • Shim, Joo-Hyun;Park, Mi-Rung;Yang, Byoung-Chul;Ko, Yeoung-Gyu;Oh, Keon-Bong;Lee, Jeong-Woong;Woo, Jae-Seok;Park, Eung-Woo;Park, Soo-Bong;Hwang, Seong-Soo
    • Reproductive and Developmental Biology
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    • v.33 no.3
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    • pp.157-162
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    • 2009
  • This study was performed to comprehend the developmental characteristics of cloned embryos knocked out (KO) of $\alpha$-1,3-galactosyltransferase (GalT) gene. Immature oocytes were collected and cultured for 40 hrs (1-step) or 20hrs (with hormone) + 20hrs (without hormone) (2-step). The embryos transferred with miniature pig ear fibroblast cell were used as control. The reconstructed embryos were cultured in PZM-3 with 5% $CO_2$ in air at $38.5^{\circ}C$ for 6 days. To determine the quality of the blstocysts, TUNEL and quantitative realtime RT-PCR were performed. The embryos were transferred to a surrogate (Landrace) at an earlier stage of the estrus cycle. The maturation rate was significantly higher in 2-step method than that of 1-step (p<0.05). The blastocyst development of GalT KO embryos was significantly lower than that of normal cloned embryos (p<0.05). The total and apoptotic cell number of GalT KO blastocysts was not different statistically from control. The relative abundance of Bax-$\alpha$/Bcl-xl ratio was significantly higher in both cloned blastocysts than that of in vivo blastocysts (p<0.05). Taken together, it can be postulated that the lower developmental potential and higher expression of apoptosis related genes in GalT KO SCNT embryos might be a cause of a low efficiency of GalT KO cloned miniature pig production.

Tumour-Derived Reg3A Educates Dendritic Cells to Promote Pancreatic Cancer Progression

  • Guo, Jie;Liao, Mengfan;Hu, Xianmin;Wang, Jun
    • Molecules and Cells
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    • v.44 no.9
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    • pp.647-657
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    • 2021
  • As a pancreatic inflammatory marker, regenerating islet-derived protein 3A (Reg3A) plays a key role in inflammation-associated pancreatic carcinogenesis by promoting cell proliferation, inhibiting apoptosis, and regulating cancer cell migration and invasion. This study aimed to reveal a novel immuno-regulatory mechanism by which Reg3A modulates tumour-promoting responses during pancreatic cancer (PC) progression. In an in vitro Transwell system that allowed the direct co-culture of human peripheral blood-derived dendritic cells (DCs) and Reg3A-overexpressing/ silenced human PC cells, PC cell-derived Reg3A was found to downregulate CD80, CD83 and CD86 expression on educated DCs, increase DC endocytic function, inhibit DC-induced T lymphocyte proliferation, reduce IL-12p70 production, and enhance IL-23 production by DCs. The positive effect of tumour-derived Reg3A-educated human DCs on PC progression was demonstrated in vivo by intraperitoneally transferring them into PC-implanted severe combined immunodeficiency (SCID) mice reconstituted with human T cells. A Reg3A-JAK2/STAT3 positive feedback loop was identified in DCs educated with Reg3A. In conclusion, as a tumour-derived factor, Reg3A acted to block the differentiation and maturation of the most important antigen-presenting cells, DCs, causing them to limit their potential anti-tumour responses, thus facilitating PC escape and progression.

The Studies on Embryo Production from Hanwoo Cows with High Offspring Meat Quality Grade and Embryo Transfer (후대의 육질등급이 우수한 한우 암소의 수정란 생산 및 이식에 관한 연구)

  • Lee, Hae-Lee;Park, Jae-Hee;Kim, Yong-Su;Kim, Jong-Gug
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.1
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    • pp.50-56
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    • 2019
  • These studies were conducted to evaluate developmental competence of follicular oocyte collected from the ovaries of Hanwoo cows with the high offspring meat quality (1++ and 1+ grade). Cumulus oocyte complexes from individual cows were matured, fertilized and cultured using protocols of in-vitro maturation (IVM), invitro fertilization (IVF) and in-vitro culture (IVC). The rates of blastocyst development from Hanwoo cows with the offspring meat quality grades of 1++ and 1+ were 18.6 and 21.2%, respectively. The rates of blastocyst development were 26.3, 20.7, 20.7, 17.2 and 31.2% from Hanwoo cows with the meat quality grades of 1++, 1+, 1, 2 and 3, respectively. Fiftyseven transferable embryos were recovered from 11 Hanwoo donor cows (5.2/head) with the high offspring meat quality grades of 1++ and 1+ in vivo, and the pregnancy rate after embryo transfer was 61.1%. In conclusion, these results suggest that in vitro embryo production from the ovaries of cows with the high meat quality grades using individual culture system can be used an efficient method for livestock improvement. In addition, for the successful industrialization of embryo transfer, conception rate should be improved.

Tissue integration patterns of non-crosslinked and crosslinked collagen membranes: an experimental in vivo study

  • Xiang Jin;Jin-Young Park;Jung-Seok Lee;Ui-Won Jung;Seong-Ho Choi;Jae-Kook Cha
    • Journal of Periodontal and Implant Science
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    • v.53 no.3
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    • pp.207-217
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    • 2023
  • Purpose: Non-crosslinked and crosslinked collagen membranes are known to exhibit distinct degradation characteristics, resulting in contrasting orientations of the adjacent tissues and different biological processes. The aim of this study was to conduct a histomorphometric assessment of non-crosslinked and crosslinked collagen membranes regarding neovascularization, tissue integration, tissue encapsulation, and biodegradation. Methods: Guided bone regeneration was performed using either a non-crosslinked (BG) or a crosslinked collagen membrane (CM) in 15 beagle dogs, which were euthanized at 4, 8, and 16 weeks (n=5 each) for histomorphometric analysis. The samples were assessed regarding neovascularization, tissue integration, encapsulation, the remaining membrane area, and pseudoperiosteum formation. The BG and CM groups were compared at different time periods using nonparametric statistical methods. Results: The remaining membrane area of CM was significantly greater than that of BG at 16 weeks; however, there were no significant differences at 4 and 8 weeks. Conversely, the neovascularization score for CM was significantly less than that for BG at 16 weeks. BG exhibited significantly greater tissue integration and encapsulation scores than CM at all time periods, apart from encapsulation at 16 weeks. Pseudoperiosteum formation was observed in the BG group at 16 weeks. Conclusions: Although BG membranes were more rapidly biodegraded than CM membranes, they were gradually replaced by connective tissue with complete integration and maturation of the surrounding tissues to form dense periosteum-like connective tissue. Further studies need to be performed to validate the barrier effect of the pseudoperiosteum.