• Title/Summary/Keyword: Technology Differentiation

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Functional Classification of Gene Expression Profiles During Differentiation of Mouse Embryonic Cells on Monolayer Culture

  • Leem, Sun-Hee;Ahn, Eun-Kyung;Heo, Jeong-Hoon
    • Animal cells and systems
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    • v.13 no.2
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    • pp.235-245
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    • 2009
  • Embryonic stem (ES) cells have a capability to generate all types of cells. However, the mechanism by which ES cells differentiate into specific cell is still unclear. Using microarray technology, the differentiation process in mouse embryonic stem cells was characterized by temporal gene expression changes of mouse ES cells during differentiation in a monolayer culture. A large number of genes were differentially regulated from 1 day to 14 days, and less number of genes were differentially expressed from 14 days to 28 days. The number of up-regulated genes was linearly increased throughout the 28 days of in vitro differentiation, while the number of down-regulated genes reached the plateau from 14 days to 28 days. Most differentially expressed genes were functionally classified into transcriptional regulation, development, extra cellular matrix (ECM),cytoskeleton organization, cytokines, receptors, RNA processing, DNA replication, chromatin assembly, proliferation and apoptosis related genes. While genes encoding ECM proteins were up-regulated, most of the genes related to proliferation, chromatin assembly, DNA replication, RNA processing, and cytoskeleton organization were down-regulated at 14 days. Genes known to be associated with embryo development or transcriptional regulation were differentially expressed mostly after 14 days of differentiation. These results indicate that the altered expression of ECM genes constitute an early event during the spontaneous differentiation, followed by the inhibition of proliferation and lineage specification. Our study might identify useful time-points for applying selective treatments for directed differentiation of mouse ES cells.

Inhibitory effects of Porphyra dentata extract on 3T3-L1 adipocyte differentiation

  • Choi, Su-Young;Lee, Su Yeon;Jang, Da hye;Lee, Suk Jun;Cho, Jeong-Yong;Kim, Sung-Hak
    • Journal of Animal Science and Technology
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    • v.62 no.6
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    • pp.854-863
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    • 2020
  • This study was aimed to investigate the inhibitory effects of Porphyra dentata (P. dentata) extract on the adipogenesis of 3T3-L1 cells and evaluate its anti-obesity effect. The proliferation of 3T3-L1 cells and differentiation of adipocytes under treatment of P. dentata extract was examined by measuring the cell viability using alamarBlue assay and lipid droplets by Oil Red O staining. Results showed that P. dentata extract has no cytotoxicity effect and lipid droplets formation decreased in a concentration-dependent manner in 3T3-L1 cells. It has been confirmed that transcription factors affecting lipid accumulation and anti-adipogenic effects during cell differentiation are linked to P. dentata extract. We observed that P. dentata shows lowering the mRNA expression of peroxisome proliferator-activated receptor γ2 (PPARγ2), CCAAT/enhancer binding protein α (C/EBPα) that adipogenesis-associated key transcription factors and inhibiting adipogenesis in the early stages of differentiation. Treating the cells with P. dentata did not only suppressed PPARγ2 and C/EBPα but also significantly decreased the mRNA expression of adiponectin, Leptin, fatty acid synthase, adipocyte protein 2, and Acetyl-coA carboxylase 1. Overall, the P. dentata extract demonstrated inhibitory property in adipogenesis, which has a potential effect in anti-obesity in 3T3-L1 cells.

Inhibitory Effects of Saururus Chinensis Extracts on Osteoclast Differentiation

  • Shim, Ki-Shuk;Kim, Soon-Nam;Kim, Myung-Hee;Kim, Young-Sup;Ryu, Shi-Yong;Min, Yong-Ki;Kim, Seong-Hwan
    • Natural Product Sciences
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    • v.14 no.2
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    • pp.113-117
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    • 2008
  • Saururus chinensis is a commonly used folk herb for the treatment of edema and liver diseases in Korea. To study the biological activity of Saururus chinensis in bone metabolism, we evaluated the effect of its extracts on osteoclast differentiation in vitro using primary mouse bone marrow-derived macrophages. Methanol extract (ME) from dried roots of Saururus chinensis was partitioned into methylene chloride (MF), ethyl acetate (EF), n-butanol (BF) and water fractions (WF). Tartrate-resistance acid phosphatase (TRAP) activity assay and western blot analysis were performed to determine the effect on osteoclast differentiation and mitogen-activated protein (MAP) kinases activation. ME, MF and EF dramatically inhibited receptor activator of ${NF-kB}$ ligand (RANKL)-induced formation of multinucleated osteoclasts and activation of MAP kinases. This study firstly demonstrated that ME, MF and EF of Saururus chinensis have the potential to inhibit the osteoclast differentiation, which results from the inhibition of MAP kinases activations in part.

Effects of Sex Steroid Hormones on Differentiation of Pig Preadipocytes (스테로이드 성호르몬이 돼지 지방전구세포의 분화에 미치는 영향)

  • Kim, H.R.;Lee, K.H.;Choi, I.H.;Chung, C.S.
    • Journal of Animal Science and Technology
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    • v.49 no.5
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    • pp.593-598
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    • 2007
  • The current study was undertaken to determine the effects of sex steroid hormones(estrogen, testosterone and 19-nortestosterone) on differentiation and proliferation of pig preadipocytes. The preadipocytes were isolated from the backfat of new-born female pigs by collagenase digestion. 10-8M and 10-7M sex steroid hormones were treated to the cultured preadipocytes. Sex steroid hormones treated during the early stage of cell growth did not affect differentiation and proliferation of preadipocytes. However, testosterone and 19- nortestosterone treated during the late stage of cell growth stimulated differentiation of pig preadipocytes.

Effect of Conjugated Linoleic Acid(CLA) on Proliferation and Differentiation of Porcine Adipocyte and Muscle Cell (Conjugated Linoleic Acid(CLA)가 돼지 지방세포와 근육세포의 증식과 분화에 미치는 영향)

  • Chung, C.S.;Kim, H.R.;Kang, J.N.;Kim, N.S.
    • Journal of Animal Science and Technology
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    • v.49 no.1
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    • pp.25-32
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    • 2007
  • The current study was undertaken to determine the effect of conjugated linoleic acid(CLA) isomers, cis-9, cis-11(c9c11), cis-9, trans-11(c9t11), trans-9, trans-11(t9t11), trans-10, cis-12(t10c12) on differentiation of pig preadipocytes and myogenic satellite cells during culture. Cells were isolated from new born pigs. The t10c12 isomer decreased differentiation of pig preadipocytes(92%), but not that of myogenic cells. The t9t11 isomer decreased differentiation of preadipocytes(14%) and increased that of myogenic cells (26%). No other CLA isomers affected differentiation of preadipocytes or myogenic cells. The effects of CLA on proliferation of preadipocytes and myogenic cells were small, compared to the effects on differentiation. These results suggest that CLA isomers have different effects on differentiaton of pig preadipocytes and myogenic cells.

Effect of Isomers of Conjugated Linoleic Acid on Porcine Preadipocyte Differentiation (Conjugated Linoleic Acid(CLA) 이성체가 돼지 지방전구세포의 분화에 미치는 영향)

  • Moon, H.S;Chung, C.S.
    • Journal of Animal Science and Technology
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    • v.46 no.6
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    • pp.967-974
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    • 2004
  • The current study was undertaken to determine the effect of various conjugated linoleic acid (CLA) isomers on differentiation of pig preadipocyte during culture. Preadipocyte(stroma-vascular cell) was isolated from the backfat of newborn pigs and cultured to differentiate into mature fat cell. Different doses of CLA isomers were treated to the culture media at different times. Cell differentiation was determined by measuring the glycerol3-phosphate dehydrogenase activity of the cultured preadipocytes. Twenty and fifty $\mu$M of trans110_cis 12 isomer of CLA inhibited differentiation of pig preadipocyte whereas cis9-cis II isomer stimulated the differentiation. Both cis9-transII and trans9-trans11 isomers showed no effect. Effect of CLA isomer was more evident at the early stage of culture(day 0-8), than the late stage(day 8-14). These results suggest that each CLA isomer has different effect on pig preadipocyte differentiation.

Microarray Profiling of Genes Differentially Expressed during Erythroid Differentiation of Murine Erythroleukemia Cells

  • Heo, Hyen Seok;Kim, Ju Hyun;Lee, Young Jin;Kim, Sung-Hyun;Cho, Yoon Shin;Kim, Chul Geun
    • Molecules and Cells
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    • v.20 no.1
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    • pp.57-68
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    • 2005
  • Murine erythroleukemia (MEL) cells are widely used to study erythroid differentiation thanks to their ability to terminally differentiate in vitro in response to chemical induction. At the molecular level, not much is known of their terminal differentiation apart from activation of adult-type globin gene expression. We examined changes in gene expression during the terminal differentiation of these cells using microarray-based technology. We identified 180 genes whose expression changed significantly during differentiation. The microarray data were analyzed by hierarchical and k-means clustering and confirmed by semi-quantitative RT-PCR. We identified several genes including H1f0, Bnip3, Mgl2, ST7L, and Cbll1 that could be useful markers for erythropoiesis. These genetic markers should be a valuable resource both as potential regulators in functional studies of erythroid differentiation, and as straightforward cell type markers.

Emodin stimulates the osteoblast differentiation via activating bone morphogenetic protein-2 gene expression at low concentration

  • Cheon, Myeong-Sook;Lee, Su-Ui;Kim, Ho-Kyoung;Kim, Young-Sup;Min, Yong-Ki;Kim, Seong-Hwan
    • Korean Journal of Oriental Medicine
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    • v.13 no.1 s.19
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    • pp.139-145
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    • 2007
  • Emodin is one of the main active components of a traditional Korean medicine isolated from the root and rhizomes of Rheum palmatum L. In this study, of 222 natural compounds to evaluate the anabolic activities, emodin activated bone morphogenetic protein (BMP)-2 promoter in the differentiation process of mouse osteoblastic MC3T3-E1 cells. Emodin was shown to significantly stimulate the activity and expression of alkaline phosphatase, an earlyphase marker of osteoblastic differentiation, on the differentiation day 7, and induce the osteopontin mRNA expression from the differentiation day 14. In addition, low concentration (up to 5 M) of emodin dramatically promoted the induction of mineralization in MC3T3-E1 subclone 4 cells. The stimulatory effect of emodin on the osteoblast differentiation/mineralization could be associated with its potential to stimulate the BMP-2 gene expression. Although further studies are needed to determine the precise mechanism, this study suggests that the use of herbal medicine containing natural compounds with anabolic activity such as emodin could have a beneficial effect on bone health.

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Relationship between Endogenous Polyamines and Floral Bud Differentiation in Chrysanthemum morifolium under Short-day Conditions

  • Guo, Jun-E;Li, Tian;Sun, Xianzhi;Zheng, Chengshu;Sun, Xia
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.31-38
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    • 2015
  • This study was carried out to investigate the relationship between endogenous polyamines (PAs) and floral bud differentiation in chrysanthemum (Chrysanthemum morifolium). In this study, PA content (free, bound, and conjugated) in apical buds, leaves, and roots changed appreciably during floral bud differentiation. PAs accumulated during series of processes such as floral induction, differentiation of floret primordia, and crown formation in apical buds; changes in PAs in apical buds may have a relationship with those in leaves and roots. The levels of free PAs and conjugated PAs [putrescine (Put) and spermine (Spm)] in apical buds rapidly increased during the initiation stage of floral bud differentiation, while free and conjugated spermidine (Spd) reached their highest levels at the stage of floret primordium differentiation. In the free, conjugated, and bound PA fractions, the changes in Spm content were negligible compared to those of Put and Spd throughout the experiment. These findings indicate that PAs participate in regulating the process of flower bud differentiation in chrysanthemum.

Conjugation of vascular endothelial growth factor to poly lactic-co-glycolic acid nanospheres enhances differentiation of embryonic stem cells to lymphatic endothelial cells

  • Yoo, Hyunjin;Choi, Dongyoon;Choi, Youngsok
    • Animal Bioscience
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    • v.34 no.4
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    • pp.533-538
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    • 2021
  • Objective: Pluripotent stem cell-derived lymphatic endothelial cells (LECs) show great promise in their therapeutic application in the field of regenerative medicine related to lymphatic vessels. We tested the approach of forced differentiation of mouse embryonal stem cells into LECs using biodegradable poly lactic-co-glycolic acid (PLGA) nanospheres in conjugation with growth factors (vascular endothelial growth factors [VEGF-A and VEGF-C]). Methods: We evaluated the practical use of heparin-conjugated PLGA nanoparticles (molecular weight ~15,000) in conjugation with VEGF-A/C, embryoid body (EB) formation, and LEC differentiation using immunofluorescence staining followed by quantification and quantitative real-time polymerase chain reaction analysis. Results: We showed that formation and differentiation of EB with VEGF-A/C-conjugated PLGA nanospheres, compared to direct supplementation of VEGF-A/C to the EB differentiation media, greatly improved yield of LYVE1(+) LECs. Our analyses revealed that the enhanced potential of LEC differentiation using VEGF-A/C-conjugated PLGA nanospheres was mediated by elevation of expression of the genes that are important for lymphatic vessel formation. Conclusion: Together, we not only established an improved protocol for LEC differentiation using PLGA nanospheres but also provided a platform technology for the mechanistic study of LEC development in mammals.