• Title/Summary/Keyword: In vitro expansion

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The effect of repeated porcelain firings on corrosion resistance of different dental alloys

  • Tuncdemir, Ali Riza;Karahan, Ismail;Polat, Serdar;Malkoc, Meral Arslan;Dalkiz, Mehmet
    • The Journal of Advanced Prosthodontics
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    • v.5 no.1
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    • pp.44-50
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    • 2013
  • PURPOSE. The aim of this study was to evaluate the effects of repeated porcelain firing process on the corrosion rates of the dental alloys. MATERIALS AND METHODS. Cr-Co, Cr-Ni and Pd-Ag alloys were used for this study. Each metal supported porcelain consisted of 30 specimens of 10 for 7, 9 and 11 firing each. Disc-shaped specimens 10 mm diameter and 3 mm thickness were formed by melting alloys with a propane-oxygen flame and casted with a centrifuge casting machine and then with the porcelain veneer fired onto the metal alloys. Corrosion tests were performed in quintuplicate for each alloy (after repeated porcelain firing) in Fusayama artificial saliva solution (pH = 5) in a low thermal-expansion borosilicate glass cell. Tamhane and Sheffe test was used to compare corrosion differences in the results after repeated firings and among 7, 9 and 11 firing for each alloy. The probability level for statistical significance was set at ${\alpha}$=0.05. RESULTS. The corrosion resistance was higher (30 mV), in case of 7 times firing (Commercial). On the other hand, it was lower in case of 11 times firing (5 mV) (P<.05). Conclusion. Repeated firings decreased corrosion resistance of Pd-Ag, Cr-Co and Cr-Ni alloys. The Pd-Ag alloy exhibited little corrosion in in vitro tests. The Cr-Ni alloy exhibited higher corrosion resistance than Cr-Co alloys in in vitro tests.

Characterization of human cardiac mesenchymal stromal cells and their extracellular vesicles comparing with human bone marrow derived mesenchymal stem cells

  • Kang, In Sook;Suh, Joowon;Lee, Mi-Ni;Lee, Chaeyoung;Jin, Jing;Lee, Changjin;Yang, Young Il;Jang, Yangsoo;Oh, Goo Taeg
    • BMB Reports
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    • v.53 no.2
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    • pp.118-123
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    • 2020
  • Cardiac regeneration with adult stem-cell (ASC) therapy is a promising field to address advanced cardiovascular diseases. In addition, extracellular vesicles (EVs) from ASCs have been implicated in acting as paracrine factors to improve cardiac functions in ASC therapy. In our work, we isolated human cardiac mesenchymal stromal cells (h-CMSCs) by means of three-dimensional organ culture (3D culture) during ex vivo expansion of cardiac tissue, to compare the functional efficacy with human bone-marrow derived mesenchymal stem cells (h-BM-MSCs), one of the actively studied ASCs. We characterized the h-CMSCs as CD90low, c-kitnegative, CD105positive phenotype and these cells express NANOG, SOX2, and GATA4. To identify the more effective type of EVs for angiogenesis among the different sources of ASCs, we isolated EVs which were derived from CMSCs with either normoxic or hypoxic condition and BM-MSCs. Our in vitro tube-formation results demonstrated that the angiogenic effects of EVs from hypoxia-treated CMSCs (CMSC-Hpx EVs) were greater than the well-known effects of EVs from BM-MSCs (BM-MSC EVs), and these were even comparable to human vascular endothelial growth factor (hVEGF), a potent angiogenic factor. Therefore, we present here that CD90lowc-kitnegativeCD105positive CMSCs under hypoxic conditions secrete functionally superior EVs for in vitro angiogenesis. Our findings will allow more insights on understanding myocardial repair.

The effects of properties and interactions of surface molecules in antigen presenting cells on T cell activation (인공 항원제시세포의 표면 분자의 특성 및 상호작용이 T 세포 활성화에 미치는 영향)

  • Min, Youngsil;Kang, Yoon Joong
    • Journal of Convergence for Information Technology
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    • v.10 no.6
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    • pp.164-176
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    • 2020
  • Efficient production of antigen specific cytotoxic T cells is critical for appropriate adoptive immune response. In vitro culture and expansion of human T lymphocyte clones are very sophisticated and subtle procedure in immune cell therapy and hard to control. Therefore, many groups devoted their efforts to manipulate artificial antigen presenting cells (aAPCs) that can induce T cell activation and clonal expansion. To mimicking of natural antigen-presenting cells, aAPCs encompass basic signal molecules required for T cell activation: MHC:antigen complexes, co-stimulatory molecules and soluble immune modulating molecules. Orchestrated organization of these molecules is important for efficient T cell activation. Here, we discuss how those molecules have been incorporated in several aAPC models, but also how physical properties od aAPC are important for interaction with T cells.

Effects of Prostaglandins on In Vitro Development of Bovine Embryos (소 체외 수정란의 체외 발육에 미치는 Prostaglandins의 영향)

  • Shin, S.O.;Park, S.B.;Park, C.K.
    • Journal of Embryo Transfer
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    • v.22 no.1
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    • pp.27-32
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    • 2007
  • This study was conducted to examine the effects of prostaglandin $F_2{\alpha}(PGF_2{\alpha})$ and prostaglandin $E_2 (PGE_2)$ on the expansion and hatching of bovine embryos. During the in vitro culture, embryos were cultured with the following groups: (1) 0, 1, 10 and 100ng/ml $PGF_2{\alpha}$ (2) 0, 1, 10 and 100 ng/ml $PGF_2{\alpha}$, (3) low concentration of $PGF_2{\alpha}$ ; low concentration of $PGF_2{\alpha}$, (1ng/ml : 1ng/ml), (4) low concentration of $PGF_2{\alpha}$ : high concentration of $PGF_2{\alpha}$ (1ng/ml : 10ng/ml) (5) high concentration of $PGF_2{\alpha}$ : low concentration of $PGE_2$ (10ng/ml 1ng/ml) (6) high concentration of $PGF_2{\alpha}$ : high concentration of $PGE_2$(10 ng/ml : 10ng/ml). In the results of this study, treatment of $PGF_2{\alpha}$ or $PGE_2$ did not affect in vitro development to blastocysts. However, the hatching rates of embryos cultured with 10ng/ml $PGE_2$(10.3%) and 1ng/ml $PGF_2{\alpha}$ 10ng/ml $PGE_2$(22.2%) were significantly (P<0.05) higher than in control (4.3% and 12.7%) and other treatment groups. All groups treated with high concentrations of $PGF_2{\alpha}$ showed decreased hatching rates. Thus, this results suggested that $PGF_2{\alpha}\;and\;PGE_2$ were concerned with the hatching in bovine embryos, and their effects on hatching were different by the concentrations.

Vitrification solution without sucrose for cryopreservation in mouse blastocysts

  • Joo, Jong Kil;Lee, Young Ju;Jeong, Ju Eun;Kim, Seung Chul;Ko, Gyoung Rae;Lee, Kyu Sup
    • Clinical and Experimental Reproductive Medicine
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    • v.41 no.3
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    • pp.115-119
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    • 2014
  • Objective: This study was designed to investigate the survival rate of vitrified mouse blastocysts depending on the presence or absence of sucrose in vitrification solution. Methods: Mouse two-cell embryos were collected and cultured to blastocysts. Two vitrification solutions were prepared. The control solution was composed of 25% glycerol, 25% ethylene glycol, and 0.5 M sucrose (G25E250.5S) containing 2.5 mL glycerol, 2.5 mL ethylene glycol, 2 mL SSS, and 0.855 g sucrose in 5 mL PB1. The experimental solution was composed of 25% glycerol and 25% ethylene glycol (G25E25) and contained 2.5 mL glycerol and 2.5 mL ethylene glycol in 5 mL PB1. Artificial shrinkage was conducted by aspirating the blastocoelic fluid using an ICSI pipette. To examine the effect of sucrose in the vitrification solution on the survival rate of mouse blastocysts, the shrunken-equilibrated blastocysts were rehydrated or vitrified after being exposed to one of the two vitrification solutions. After exposure and the vitrification-thawing process, the re-expansion rate and hatching rate were evaluated after 6 hours of in vitro culture. Results: The re-expansion rate of mouse blastocysts exposed to vitrification solution with and without sucrose were not different in the experimental solution (without sucrose) (98%) and the control solution (with sucrose) (92%) (p>0.05). The hatching rate was higher in the experimental solution (95%) than in the control solution (88%), but did not differ across two treatments (p>0.05). The re-expansion rate of mouse blastocysts vitrified in the control solution was 92% and 94%, respectively (p>0.05), and the hatching rate was higher in the experimental solution (90%) than in the control solution (74%) (p<0.05). Conclusion: Sucrose need not be added in vitrification solution for freezing of artificially shrunken mouse blastocysts.

CD43 Expression Regulated by IL-12 Signaling Is Associated with Survival of CD8 T Cells

  • Lee, Jee-Boong;Chang, Jun
    • IMMUNE NETWORK
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    • v.10 no.5
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    • pp.153-163
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    • 2010
  • Background: In addition to TCR and costimulatory signals, cytokine signals are required for the differentiation of activated CD8 T cells into memory T cells and their survival. Previously, we have shown that IL-12 priming during initial antigenic stimulation significantly enhanced the survival of activated CD8 T cells and increased the memory cell population. In the present study, we analyzed the mechanisms by which IL-12 priming contributes to activation and survival of CD8 T cells. Methods: We observed dramatically decreased expression of CD43 in activated CD8 T cells by IL-12 priming. We purified $CD43^{lo}$ and $CD43^{hi}$ cells after IL-12 priming and analyzed the function and survival of each population both in vivo and in vitro. Results: Compared to $CD43^{hi}$ effector cells, $CD43^{lo}$ effector CD8 T cells exhibited reduced cytolytic activity and lower granzyme B expression but showed increased survival. $CD43^{lo}$ effector CD8 T cells also showed increased in vivo expansion after adoptive transfer and antigen challenge. The enhanced survival of $CD43^{lo}$ CD8 T cells was also partly associated with CD62L expression. Conclusion: We suggest that CD43 expression regulated by IL-12 priming plays an important role in differentiation and survival of CD8 T cells.

NCAM as a cystogenesis marker gene of PKD2 overexpression

  • Yoo, Kyung-Hyun;Lee, Tae-Young;Yang, Moon-Hee;Park, Eun-Young;Yook, Yeon-Joo;Lee, Hyo-Soo;Park, Jong-Hoon
    • BMB Reports
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    • v.41 no.8
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    • pp.593-596
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    • 2008
  • ADPKD (Autosomal Dominant Polycystic Kidney Disease) is characterized by the progressive expansion of multiple cystic lesions in the kidneys. ADPKD is caused by mutations in Ed-pl. consider PKD1 and PKD2. Recently a relation between c-myc and the pathogenesis of ADPKD was reported. In addition, c-Myc is a downstream effector of PKD1. To identify the gene regulated by PKD2 and c-Myc, we performed gene expression profiling in PKD2 and c-Myc overexpressing cells using a human 8K cDNA microarray. NCAM (neuronal cell adhesion molecule) levels were significantly reduced in PKD2 overexpressing systems in vitro and in vivo. These results suggest that NCAM is an important molecule in the cystogenesis induced by PKD2 overexpession.

Development of Zygotic Embryos and Seedlings is Affected by Radiation Spectral Compositions from Light Emitting Diode (LED) System in Chestnut (Castanea crenata S. et Z.)

  • Park, So-Young;Kim, Man-Jo
    • Journal of Korean Society of Forest Science
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    • v.99 no.5
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    • pp.750-754
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    • 2010
  • Among the environmental conditions employed in micropropagation, light quality plays an important role in growth, specially morphogenesis and photosynthesis. The effect of radiation quality (350-740 nm) on the development and growth of zygotic embryos and in vitro plantlets of open-pollinated chestnut (Castanea crenata S. et Z.) were studied. Two types of explants were exposed for 4 weeks to cool white (W, as control), monochromatic red (R, peak emission 650 nm), monochromatic blue (B, peak emission 440 nm), red+blue (R+B, 1:1), or red+far-red (R+Fr, 1:1, far-red peak emission 720 nm) radiation from a light-emitting-diode (LED) system. While the zygotic embryos showed positive photoblastic behavior, their germination was inhibited by blue radiation. Hypocotyl elongation and root development were promoted by red radiation. The emergence of primary leaf and its expansion were faster under blue than under red radiation. In the plantlets, red and red+far-red radiation significantly increased the formation and growth of the root, whereas blue light reduced rooting. Therefore, radiation quality appears to influence some steps in the development of zygotic embryos and plantlets in the chestnut.

Phloroglucinol Inhibits the in vitro Differentiation Potential of CD34 Positive Cells into Endothelial Progenitor Cells

  • Kwon, Yi-Hong;Lee, Jun-Hee;Jung, Seok-Yun;Kim, Jae-Won;Lee, Sang-Hun;Lee, Dong-Hyung;Lee, Kyu-Sup;Lee, Boo-Yong;Kwon, Sang-Mo
    • Biomolecules & Therapeutics
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    • v.20 no.2
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    • pp.158-164
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    • 2012
  • Inhibiting the bioactivities of circulating endothelial progenitor cells (EPCs) results in significant inhibition of neovessel formation during tumor angiogenesis. To investigate the potential effect of phloroglucinol as an EPC inhibitor, we performed several in vitro functional assays using $CD34^+$ cells isolated from human umbilical cord blood (HUCB). Although a high treatment dose of phloroglucinol did not show any cell toxicity, it specifically induced the cell death of EPCs under serum free conditions through apoptosis. In the EPC colony-forming assay (EPC-CFA), we observed a significant decreased in the small EPC-CFUs for the phloroglucinol group, implying that phloroglucinol inhibited the early stage of EPC commitment. In addition, in the in vitro expansion assay using $CD34^+$ cells, treatment with phloroglucinol was shown to inhibit endothelial lineage commitment, as demonstrated by the decrease in endothelial surface markers of EPCs including $CD34^+$, $CD34^+/CD133^+$, $CD34^+/CD31^+$ and $CD34^+/CXCR4^+$. This is the first report to demonstrate that phloroglucinol can inhibit the functional bioactivities of EPCs, indicating that phloroglucinol may be used as an EPC inhibitor in the development of biosafe anti-tumor drugs that target tumor angiogenesis.

The Kinetics of Secondary Response of Antigen-Specific $CD4^+$ T Cells Primed in vitro with Antigen (실험적으로 항원에 의하여 일차 자극된 $CD4^+$ T 세포의 이차 면역 반응의 분석)

  • Park, Seong-Ok;Han, Young-Woo;Aleyas, Abi George;George, June Abi;Yoon, Hyun-A;Eo, Seong-Kug
    • IMMUNE NETWORK
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    • v.6 no.2
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    • pp.93-101
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    • 2006
  • Background: Memory T lymphocytes of the immune system provide long-term protection in response to bacterial or viral infections/immunization. Ag concentration has also been postulated to be important in determining whether T cell differentiation favors effector versus memory cell development. In the present study we hypothesized that naive Ag-specific $CD4^+$ T cells briefly stimulated with different Ag doses at the primary exposure could affect establishment of memory cell pool after secondary immunization. Methods: To assess this hypothesis, the response kinetics of DO11.10 TCR $CD4^+$ T cells primed with different Ag doses in vitro was measured after adoptive transfer to naive BALB/c mice. Results: Maximum expansion was shown in cells primarily stimulated with high doses of ovalbumin peptide $(OVA_{323-339})$, whereas cells in vitro stimulated with low dose were expanded slightly after in vivo secondary exposure. However, the cells primed with low $OVA_{323-339}$ peptide dose showed least contraction and established higher number of memory cells than other treated groups. When the cell division was analyzed after adoptive transfer, the high dose Ag-stimulated donor cells have undergone seven rounds of cell division at 3 days post-adoptive transfer. However, there was very few division in naive and low dose of peptide-treated group. Conclusion: These results suggest that primary stimulation with a low dose of Ag leads to better memory $CD4^+$ T cell generation after secondary immunization. Therefore, these facts imply that optimally primed $CD4^+$ T cells is necessary to support effective memory pool following administration of booster dose in prime-boost vaccination.