• Title/Summary/Keyword: Cell formation

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Establishment of In-Vitro Culture System for Enhancing Production of Somatic Cell Nuclear Transfer (SCNT) Blastocysts with High Performance in the Colony Formation and Formation of Colonies Derived from SCNT Blastocysts in Pigs

  • Han, Na Rae;Baek, Song;Lee, Yongjin;Lee, Joohyeong;Yun, Jung Im;Lee, Eunsong;Lee, Seung Tae
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.2
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    • pp.130-138
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    • 2019
  • Although somatic cell nuclear transfer (SCNT)-derived embryonic stem cells (ESCs) in pigs have great potential, their use is limited because the establishment efficiency of ESCs is extremely low. Accordingly, we tried to develop in-vitro culture system stimulating production of SCNT blastocysts with high performance in the colony formation and formation of colonies derived from SCNT blastocysts for enhancing production efficiency of porcine ESCs. For these, SCNT blastocysts produced in various types of embryo culture medium were cultured in different ESC culture medium and optimal culture medium was determined by comparing colony formation efficiency. As the results, ICM of porcine SCNT blastocysts produced through sequential culture of porcine SCNT embryos in the modified porcine zygote medium (PZM)-5 and the PZM-5F showed the best formation efficiency of colonies in α-MEM-based medium. In conclusion, appropriate combination of the embryo culture medium and ESC culture medium will greatly contribute to successful establishment of ESCs derived from SCNT embryos.

High Frequency Bulblet Formation in Scale and Stem Thin Cell Layer Explant Cultures of Lilium Oriental Hybrids (오리엔탈 백합의 인편 및 줄기의 박판 세포층 절편으로부터 고빈도 자구형성)

  • Oh, Seung-Cheol;Chung, Myung-Hee;Kim, Suk-Won;Liu, Jang-Ryol
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.251-255
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    • 2003
  • An efficient system for in vitro bulblet formation of Lilium oriental hybrids(cvs). Casa Blanca and Siberia is described. Transverse thin cell layer(tTCL)(1mm thick) explants of 'Casa Blanca' formed bulblets at a frequency of 97.7% when cultured on Murashige and Skoog (MS) medium supplemented with 1mg/L 2,4 dichlorophenoxyacetic acid(2,4-D) (On average 15.6 bulblets were formed per explant). The frequency of bulblet formation was drastically reduced when the explant ghickness was thinner than 1 mm. Explants from the outermost layer of bulb scale produced greater frequency of bulblet formation than middle or innermost layer. Among auxins supplemented to culture medium at 1 mg/L, 2,4-D led to greater frequency of bulblet formation on explants than dicamba, picamba, or phenylacetic acid(PAA). tTCL explants from the middle region of the outermost layer bulb scale yielded greater frequency of bulblet formation than the upper or lower region. tTCL stem explants of 'Siberia' formed bulblets at a frequency of 95.3% when cultured on MS medium with 1 mg/L 2,4-D(On average 9.1 bulblets were formed per explant). The system estabilished in this study will be useful for in vitro rapid propagation and genetic transformation of Lilium Oriental hybrids.

Suspension Culture-Mediated Tetraploid Formation in Mouse Embryonic Stem Cells

  • Lee, Jae-Hee;Gong, Seung-Pyo;Lim, Jeong-Mook;Lee, Seung-Tae
    • Reproductive and Developmental Biology
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    • v.36 no.1
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    • pp.21-26
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    • 2012
  • Suspension culture is a useful tool for culturing embryonic stem (ES) cells in large-scale, but the stability of pluripotency and karyotype has to be maintained $in$ $vitro$ for clinical application. Therefore, we investigated whether the chromosomal abnormality of ES cells was induced in suspension culture or not. The ES cells were cultured in suspension as a form of aggregate with or without mouse embryonic fibroblasts (MEFs), and 0 or 1,000 U/ml leukemia inhibitory factor (LIF) was treated to suspended ES cells. After culturing ES cells in suspension, their karyotype, DNA content, and properties of pluripotency and differentiation were evaluated. As a result, the formation of tetraploid ES cell population was significantly increased in suspension culture in which ES cells were co-cultured with both MEFs and LIF. Tetraploid ES cell population was also generated when ES cells were cultured alone in suspension regardless of the existence of LIF. On the other hand, the formation of tetraploid ES cell population was not detected in LIF-free condition, in which MEFs were included. The origin of tetraploid ES cell population was turned out to be E14 ES cells and not MEFs by microsatellite analysis and the basic properties of them were still maintained despite ploidy-conversion to tetraploidy. Furthermore, we identified the ploidy shift from tetraploidy to near-triploidy as tetraploid ES cells were differentiated spontaneously. From these results, we demonstrated that suspension culture system could induce ploidy-conversion generating tetraploid ES cell population. Moreover, optimization of suspension culture system may make possible mass-production of ES cells.

Production of Bacillus thuringiensis Spore Using an Industrial Medium (산업용 배지를 이용한 Bacillus thuringiensis의 포지생산)

  • 최성호;강석권;유연우
    • KSBB Journal
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    • v.13 no.6
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    • pp.644-648
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    • 1998
  • In the production of a low cost bacterial insecticide, it is important to produce a high spore concentration using low price substrates. Experiments were carried out to investigate the effects of the addition of mineral salts and glucose, and of dissolved oxygen concentration on the cell growth and spore formation of Bacillus thuringiensis var aizawai using a cheap wheat and soybean meal in the batch culture. The maximum viable cell number was 1.2${\times}$109 CFU/mL at 12 hr culture and spore yield was 54.2% at 74 hr culture using an industrial medium containing 20 g/L wheat meal and 30 g/L soybean meal under 1.0 vvm aeration and 200 rpm agitation. The cell growth and the spore formation were not enhanced by the addition of mineral salts in industrial medium, whereas th addition of 10g/L glucose decreased the cell growth and spore formation. We could obtain a maximum viable cell number of 2.2${\times}$109 CFU/mL and spore number of 1.9${\times}$109 CFU/mL at the dissolved oxygen concentration of 60% of saturation. The spore concentration was enhanced approximately by 2 times as compared to the dissolved oxygen concentration of 50%. In the bench-scale culture, the maximum viable cell and spore number were 2.5${\times}$109 CFU/mL, and 2.2${\times}$109 CFU/mL, respectively under 1.0 vvm aeration and 400 rpm agitation. The spore yield was 88% based on the maximum viable cell number. As a result, it was confirmed that the production of high spore concentration could be obtained by a bench-scale culture using an industrial medium.

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Genome-Wide Analysis Identifies NURR1-Controlled Network of New Synapse Formation and Cell Cycle Arrest in Human Neural Stem Cells

  • Kim, Soo Min;Cho, Soo Young;Kim, Min Woong;Roh, Seung Ryul;Shin, Hee Sun;Suh, Young Ho;Geum, Dongho;Lee, Myung Ae
    • Molecules and Cells
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    • v.43 no.6
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    • pp.551-571
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    • 2020
  • Nuclear receptor-related 1 (Nurr1) protein has been identified as an obligatory transcription factor in midbrain dopaminergic neurogenesis, but the global set of human NURR1 target genes remains unexplored. Here, we identified direct gene targets of NURR1 by analyzing genome-wide differential expression of NURR1 together with NURR1 consensus sites in three human neural stem cell (hNSC) lines. Microarray data were validated by quantitative PCR in hNSCs and mouse embryonic brains and through comparison to published human data, including genome-wide association study hits and the BioGPS gene expression atlas. Our analysis identified ~40 NURR1 direct target genes, many of them involved in essential protein modules such as synapse formation, neuronal cell migration during brain development, and cell cycle progression and DNA replication. Specifically, expression of genes related to synapse formation and neuronal cell migration correlated tightly with NURR1 expression, whereas cell cycle progression correlated negatively with it, precisely recapitulating midbrain dopaminergic development. Overall, this systematic examination of NURR1-controlled regulatory networks provides important insights into this protein's biological functions in dopamine-based neurogenesis.

A Cell Formation Model For Manufacturing System with Sequence Dependence (가공순서에 종속적인 제조시스템을 위한 군형성모형)

  • 홍상우
    • Journal of Korean Society of Industrial and Systems Engineering
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    • v.22 no.49
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    • pp.59-65
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    • 1999
  • This paper considers the cell formation in a manufacturing environment where the setup times and costs are significantly dependent on the sequence. The trade-off between saving on the setup costs and additional investment on new machines is considered for determining the economic number of cells. Accordingly, This paper develops a mixed integer program and mentions a variety of manufacturing situations where this model can be useful. This paper also includes an illustrative example.

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Two phase p-median approach to nondegenerate GT cell formation (GT 생산시스템에서 비퇴화 셀 형성을 위한 2 단계 p-median 접근법)

  • 원유경
    • Proceedings of the Korean Operations and Management Science Society Conference
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    • 2001.10a
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    • pp.21-24
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    • 2001
  • This study is concerned with the development of efficient p-median approach to nondegenerate cell formation(CF) in group technology(GT) manufacturing. Unlike most of existing CF methodologies allowing degenerate cells or families that contains no parts or machines, this study attempts to find cell configuration where each machine cell contains at least two or more machines processing at least two or more parts so as to fully utilize the similarity in designing and processing parts. Nondegenerate CF seeks to minimize both the exceptional elements outside the diagonal block and the voids within the diagonal block. To find nondegenerate cells, a two-phase p-median methodology is proposed. In phase 1, the classical p-median model is implemented to find initial cells. In phase 2, bottleneck machines and parts are reassigned until no further degenerate cells and families are found. Test results on moderately medium-sized CF problems show the substantial efficiency of the proposed approach.

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Regulation of Early Steps of Chondrogenesis in the Developing Limb

  • Kang, Shin-Sung
    • Animal cells and systems
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    • v.12 no.1
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    • pp.1-9
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    • 2008
  • In the developing limb, chondrogenesis is an important prerequisite for the formation of cartilage whose template is required for bone formation. Chondrogenesis is a tightly regulated multi-step process, including mesenchymal cell recruitment/migration, prechondrogenic condensation of the mesenchymal cells, commitment to the chondrogenic lineage, and differentiation into chondrocytes. This process is controlled exquisitely by cellular interactions with the surrounding matrix and regulating factors that initiate or suppress cellular signaling pathways and transcription of specific genes in a temporal-spatial manner. Understanding the cellular and molecular mechanisms of chondrogenesis is important not only in the context of establishing basic principle of developmental biology but also in providing research direction toward preventive and/or regenerative medicine. Here, I will overview the current understanding of cellular and molecular mechanisms contributing to prechondrogenic condensation processes, the crucial steps for chondrogenesis, focusing on cell-cell and cell-matrix interactions.

Efficient Ρ-median approach to GT cell formation (GT 셀 형성을 위한 효율적 Ρ-median 접근법)

  • Won, Youkyung
    • Proceedings of the Korean Operations and Management Science Society Conference
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    • 2000.04a
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    • pp.40-43
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    • 2000
  • This paper is concerned with development of an efficient Ρ-median approach applicable to large cell formation(CF) problems. A two-phase methodology that seeks to minimize the number of exceptional elements is proposed. In phase I, two efficient Ρ-median formulations which contain fewer binary variables than existing Ρ-median formulations are constructed. These make it possible to implement large CF problem within reasonable computer runtime with commercially available linear integer programming codes. Given the initial cell configuration found with the new p-median formulations, in phase II bottleneck machines and parts are reassigned to reduce the number of exceptional elements. This procedure has the flexibility to provide the cell designer with alternative solutions. Test results on large CF problems show a substantial efficiency of the new Ρ-median formulations.

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A Possible Physiological Role of Caspase-11 During Germinal Center Reaction

  • Kang, Shin-Jung
    • Animal cells and systems
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    • v.12 no.3
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    • pp.127-136
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    • 2008
  • Caspase-11 has been known as a dual regulator of cytokine maturation and apoptosis. Although the role of caspase-11 under pathological conditions has been well documented, its physiological role has not been studied much. In the present study, we investigated a possible physiological function of caspase-11 during immune response. In the absence of caspase-11, immunized spleen displayed increased cellularity and abnormal germinal center structure with disrupted microarchitecture. The rate of cell proliferation and apoptosis in the immunized spleen was not changed in the caspase-11-deficient mice. Furthermore, the caspase-11-deficient peritoneal macrophages showed normal phagocytotic activity. However, caspase-11-/-splenocytes and macrophages showed defective migrating capacity. The dysregulation of cell migration did not seem to be mediated by caspase-3, interleukin-$1{\alpha}$ or interleukin-$1{\beta}$ which acts downstream of caspase-11. These results suggest that a direct regulation of immune cell migration by caspase-11 is critical for the formation of germinal center microarchitecture during immune response. However, humoral immunity in the caspase-11-deficient mice was normal, suggesting the formation of germinal center structure is not essential for the affinity maturation of the antibodies.