• Title/Summary/Keyword: multiple stem

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Rapid Micropropagation of Hovenia dulcis Thunb. Through in vitro Stem Nodal Cultures

  • Park, Dong-Jin;Kang, Young-Min;Jung, Ha-Na;Min, Ji-Yun;Kim, Yong-Duck;Karigar, Chandrakant S.;Choi, Myung-Suk
    • Journal of Korean Society of Forest Science
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    • v.95 no.2
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    • pp.155-159
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    • 2006
  • An efficient method for in vitro propagation of the medicinal plant Hovenia duleis, was established. Plantlets for micropropagation of H. dulcis were obtained from in vitro germinated seeds. The effectiveness of various levels of cytokinins (BAP, Kinetin and TDZ) on multiple shoot formation from stem nodes was tested. BAP (1.0 mg/L) treatment induced highest number of multiple shoots. The growth pattern of plantlet on various culture media was undertaken. The shoot elongation was optimal on 2MS basal medium without growth regulators. The in vitro rooting ability of H. dulcis shoots was examined with two-auxins IAA and IBA. The IAA (1.0 mg/L) treatments induced earliest rooting with maximum number of roots and root growth. Rooted shoots were transferred directly to small pots with artificial soil and such established plant exhibited a normal growth pattern similar to wild plantlet.

In vitro Callus formation and Plant Regeneration of Epimedium koreanum Nakai

  • Yu, Chang-Yeon;Kim, Na-Young;Kang, Won-Hee;Kim, Myong-Jo;Heo, Kwon;Choi, Won-Cheol;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.4
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    • pp.259-262
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    • 2002
  • The efficiency of regeneration of callus and explants from leaf and stem disks of Epimedium koreanum was examined on the MS media containing 2,4-D, NAA, Kinetin, BA and TDZ. Calli were formed on the 2mg/l 2,4-D media at the rate of 32% from leaf discs and 52% from stems. No callus was produced on the media which are containing BA or TDZ alone. The combination of 2,4-D and BA showed the effect on the formation of callus. The combination of 2mg/l 2,4-D and 0.lmg/l BA in the MS media had produced the highest percentage of callus formation, 50% from leaf discs and 40% from stems, respectively. The combination of 2mg/l 2,4-D and 1mg/1 BA in the MS media had affected the formation of callus in the rate of 40% from leaf discs and 25% from stems. The combined plant growth regulators of 2,4-D and BA increased the formation of calli from leaf discs, but single treatment of 2,4-D showed the highest callus formation from stems. Multiple shoots from leaf discs were formed on the media containing NAA, BA, kinetin, and TDZ. The highest number of multiple shoots were obtained 0.1mg/l NAA combined with 1mg/l kinetin. As a result, leaf discs or stems can be used for the mass propagation of Epimedium koreanum, but stem elongation of shoots from calli was not easy.

Alpha-Synuclein Inclusion Formation in Human Oligodendrocytes

  • Yoon, Ye-Seul;Ahn, Woo Jung;Ricarte, Diadem;Ortiz, Darlene;Shin, Chan Young;Lee, Seung-Jae;Lee, He-Jin
    • Biomolecules & Therapeutics
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    • v.29 no.1
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    • pp.83-89
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    • 2021
  • Multiple system atrophy (MSA) is a neurodegenerative disease characterized by presence of α-synuclein-positive inclusions in the cytoplasm of oligodendrocytes. These glial cytoplasmic inclusions (GCIs) are considered an integral part of the pathogenesis of MSA, leading to demyelination and neuronal demise. What is most puzzling in the research fields of GCIs is the origin of α-synuclein aggregates in GCIs, since adult oligodendrocytes do not express high levels of α-synuclein. The most recent leading hypothesis is that GCIs form via transfer and accumulation of α-synuclein from neurons to oligodendrocytes. However, studies regarding this subject are limited due to the absence of proper human cell models, to demonstrate the entry and accumulation of neuronal α-synuclein in human oligodendrocytes. Here, we generated mature human oligodendrocytes that can take up neuronderived α-synuclein and form GCI-like inclusions. Mature human oligodendrocytes are derived from neural stem cells via "oligosphere" formation and then into oligodendrocytes, treating the cells with the proper differentiation factors at each step. In the final cell preparations, oligodendrocytes consist of the majority population, while some astrocytes and unidentified stem cell-like cells were present as well. When these cells were exposed to α-synuclein proteins secreted from neuron-like human neuroblastoma cells, oligodendrocytes developed perinuclear inclusion bodies with α-synuclein immunoreactivity, resembling GCIs, while the stem cell-like cells showed α-synuclein-positive, scattered puncta in the cytoplasm. In conclusion, we have established a human oligodendrocyte model for the study of GCI formation, and the characterization and use of this model might pave the way for understanding the pathogenesis of MSA.

Parthenogenetic Activation of Porcine Oocytes and Isolation of Embryonic Stem Cells-like Derived from Parthenogenetic Blastocysts

  • Xu, X.M.;Hua, J.L.;Jia, W.W.;Huang, W.;Yang, C.R.;Dou, Z.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.10
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    • pp.1510-1516
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    • 2007
  • These experiments were carried out to optimize the parameters of electrical activation, methods of parthenogenetic activation and embryo culture in vitro and meanwhile to isolate embryonic stem cells-like (ESCs) derived from porcine parthenogenetic blastocysts (pPBs). These results showed that, as the electric field strength increased from 1.0 to 2.7 kV/cm, the cleavage rate of parthenogenetic embryos increased gradually but the rate of oocyte lysis was significantly increased when using 2.7 kV/cm field strength. The rate of cleavage in 2.2 and 2.7 kV/cm groups was significantly increased in comparison with that of the 1.0 kV/cm group. A voltage field strength of 2.2 kV/cm DC was used to investigate blastocyst development following activation with a single pulse of 30 or $60-{\mu}sec$ pulse duration. The optimum pulse duration was 30-${\mu}sec$, with a blastocyst rate of 20.7%. Multiple pulses were inferior to a single pulse for blastocyst yield (8.0% vs. 29.9) (p<0.05). For porcine oocyte parthenogenetic activation methods, the rates of cleavage (79.0% vs. 59.8%) and blastocysts (19.4% vs. 3.4%) were significantly increased in electrical activation in contrast to chemical activation with ionomycin/6-DMAP (p<0.05). Rates of cleavage and blastocyst formation in NCSU-23 and PZM-3 embryo media were higher than those of G1.3/G2.3 serial culture media, but there was no significant difference among the three groups. The total cell number of blastocysts in PZM-3 embryo culture media containing $5{\mu}g/ml$ insulin was significantly higher than that of the control (no insulin) ($44.3{\pm}9.1$ vs. $33.9{\pm}11.7$). For isolation of PESCs-like, the rates of porcine blastocysts attached to feeder layers and ICM colony formation in Method B (nude embryo culture) were better than those in Method A (intact embryo culture).

Enhanced Healing of Rat Calvarial Bone Defects with Hypoxic Conditioned Medium from Mesenchymal Stem Cells through Increased Endogenous Stem Cell Migration via Regulation of ICAM-1 Targeted-microRNA-221

  • Chang, Woochul;Kim, Ran;Park, Sang In;Jung, Yu Jin;Ham, Onju;Lee, Jihyun;Kim, Ji Hyeong;Oh, Sekyung;Lee, Min Young;Kim, Jongmin;Park, Moon-Seo;Chung, Yong-An;Hwang, Ki-Chul;Maeng, Lee-So
    • Molecules and Cells
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    • v.38 no.7
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    • pp.643-650
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    • 2015
  • The use of conditioned medium from mesenchymal stem cells may be a feasible approach for regeneration of bone defects through secretion of various components of mesenchymal stem cells such as cytokines, chemokines, and growth factors. Mesenchymal stem cells secrete and accumulate multiple factors in conditioned medium under specific physiological conditions. In this study, we investigated whether the conditioned medium collected under hypoxic condition could effectively influence bone regeneration through enhanced migration and adhesion of endogenous mesenchymal stem cells. Cell migration and adhesion abilities were increased through overexpression of intercellular adhesion molecule-1 in hypoxic conditioned medium treated group. Intercellular adhesion molecule-1 was upregulated by microRNA-221 in mesenchymal stem cells because microRNAs are key regulators of various biological functions via gene expression. To investigate the effects in vivo, evaluation of bone regeneration by computed tomography and histological assays revealed that osteogenesis was enhanced in the hypoxic conditioned medium group relative to the other groups. These results suggest that behavioral changes of endogenous mesenchymal stem cells through microRNA-221 targeted-intercellular adhesion molecule-1 expression under hypoxic conditions may be a potential treatment for patients with bone defects.

Expression Analyses Revealed Thymic Stromal Co-Transporter/Slc46A2 Is in Stem Cell Populations and Is a Putative Tumor Suppressor

  • Kim, Ki Yeon;Lee, Gwanghee;Yoon, Minsang;Cho, Eun Hye;Park, Chan-Sik;Kim, Moon Gyo
    • Molecules and Cells
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    • v.38 no.6
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    • pp.548-561
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    • 2015
  • By combining conventional single cell analysis with flow cytometry and public database searches with bioinformatics tools, we extended the expression profiling of thymic stromal cotransporter (TSCOT), Slc46A2/Ly110, that was shown to be expressed in bipotent precursor and cortical thymic epithelial cells. Genome scale analysis verified TSCOT expression in thymic tissue- and cell type- specific fashion and is also expressed in some other epithelial tissues including skin and lung. Coexpression profiling with genes, Foxn1 and Hoxa3, revealed the role of TSCOT during the organogenesis. TSCOT expression was detected in all thymic epithelial cells (TECs), but not in the $CD31^+$endothelial cell lineage in fetal thymus. In addition, ABC transporter-dependent side population and Sca-$1^+$ fetal TEC populations both contain TSCOT-expressing cells, indicating TEC stem cells express TSCOT. TSCOT expression was identified as early as in differentiating embryonic stem cells. TSCOT expression is not under the control of Foxn1 since TSCOT is present in the thymic rudiment of nude mice. By searching variations in the expression levels, TSCOT is positively associated with Grhl3 and Irf6. Cytokines such as IL1b, IL22 and IL24 are the potential regulators of the TSCOT expression. Surprisingly, we found TSCOT expression in the lung is diminished in lung cancers, suggesting TSCOT may be involved in the suppression of lung tumor development. Based on these results, a model for TEC differentiation from the stem cells was proposed in context of multiple epithelial organ formation.

Caenorhabditis elegans: A Model System for Anti-Cancer Drug Discovery and Therapeutic Target Identification

  • Kobet, Robert A.;Pan, Xiaoping;Zhang, Baohong;Pak, Stephen C.;Asch, Adam S.;Lee, Myon-Hee
    • Biomolecules & Therapeutics
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    • v.22 no.5
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    • pp.371-383
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    • 2014
  • The nematode Caenorhabditis elegans (C. elegans) offers a unique opportunity for biological and basic medical researches due to its genetic tractability and well-defined developmental lineage. It also provides an exceptional model for genetic, molecular, and cellular analysis of human disease-related genes. Recently, C. elegans has been used as an ideal model for the identification and functional analysis of drugs (or small-molecules) in vivo. In this review, we describe conserved oncogenic signaling pathways (Wnt, Notch, and Ras) and their potential roles in the development of cancer stem cells. During C. elegans germline development, these signaling pathways regulate multiple cellular processes such as germline stem cell niche specification, germline stem cell maintenance, and germ cell fate specification. Therefore, the aberrant regulations of these signaling pathways can cause either loss of germline stem cells or overproliferation of a specific cell type, resulting in sterility. This sterility phenotype allows us to identify drugs that can modulate the oncogenic signaling pathways directly or indirectly through a high-throughput screening. Current in vivo or in vitro screening methods are largely focused on the specific core signaling components. However, this phenotype-based screening will identify drugs that possibly target upstream or downstream of core signaling pathways as well as exclude toxic effects. Although phenotype-based drug screening is ideal, the identification of drug targets is a major challenge. We here introduce a new technique, called Drug Affinity Responsive Target Stability (DARTS). This innovative method is able to identify the target of the identified drug. Importantly, signaling pathways and their regulators in C. elegans are highly conserved in most vertebrates, including humans. Therefore, C. elegans will provide a great opportunity to identify therapeutic drugs and their targets, as well as to understand mechanisms underlying the formation of cancer.

Occurrence of Tiller and Its Effects on Grain Yield of Barley under Different Plant Densities. (대맥의 파종밀도가 분얼발생 및 각얼자의 특성과 수량에 미치는 영향)

  • 신만균;맹돈재;하용웅
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.32 no.3
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    • pp.294-301
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    • 1987
  • This experiment was carried out to investigate the occurrence of tiller and its proportion to grain yield under 5 different plant densities. Plant density 5 x 5cm was more favorable for increasing the number of spikes per unit area, where leads to improve grain yield. Effective tillers showed only main stem, M$_1$-axil and M$_2$-axil, indication more effective tillers in M$_1$ rather than main stem. Every tillers held at least 6 leaves to bear spikes, indicating that main stem has 12 -13 leaves. Leaves attached on main stem M$\sub$0/ and M$_1$ were very important as main source of photosynthesis, especially under more compart plant densities while leaves of M$_1$ occupied 66.8% in a plant. The proportion of grain yield of each tiller to total grain yield per plant was high in main stem under conpact plant density, in M$_1$ stems under less compact plant density, and in M$_2$ -stems under wide plant density, indicating same result on multiple regression analysis.

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Single cell heterogeneity in human pluripotent stem cells

  • Yang, Seungbok;Cho, Yoonjae;Jang, Jiwon
    • BMB Reports
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    • v.54 no.10
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    • pp.505-515
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    • 2021
  • Human pluripotent stem cells (hPSCs) include human embryonic stem cells (hESCs) derived from blastocysts and human induced pluripotent stem cells (hiPSCs) generated from somatic cell reprogramming. Due to their self-renewal ability and pluripotent differentiation potential, hPSCs serve as an excellent experimental platform for human development, disease modeling, drug screening, and cell therapy. Traditionally, hPSCs were considered to form a homogenous population. However, recent advances in single cell technologies revealed a high degree of variability between individual cells within a hPSC population. Different types of heterogeneity can arise by genetic and epigenetic abnormalities associated with long-term in vitro culture and somatic cell reprogramming. These variations initially appear in a rare population of cells. However, some cancer-related variations can confer growth advantages to the affected cells and alter cellular phenotypes, which raises significant concerns in hPSC applications. In contrast, other types of heterogeneity are related to intrinsic features of hPSCs such as asynchronous cell cycle and spatial asymmetry in cell adhesion. A growing body of evidence suggests that hPSCs exploit the intrinsic heterogeneity to produce multiple lineages during differentiation. This idea offers a new concept of pluripotency with single cell heterogeneity as an integral element. Collectively, single cell heterogeneity is Janus-faced in hPSC function and application. Harmful heterogeneity has to be minimized by improving culture conditions and screening methods. However, other heterogeneity that is integral for pluripotency can be utilized to control hPSC proliferation and differentiation.

Health-related Quality of Life of Children and Adolescents after Hematopoietic Stem Cell Transplantation

  • Hwang, Yu Min;Bang, Kyung-Sook
    • Child Health Nursing Research
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    • v.26 no.4
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    • pp.402-410
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    • 2020
  • Purpose: This study aimed to determine the level of health-related quality of life (HRQoL) of children and adolescents who received hematopoietic stem cell transplantation (HSCT) and to examine factors influencing HRQoL. Methods: This cross-sectional descriptive study involved 85 participants aged 10 to 19 years who received treatment from 3 months to 5 years after HSCT. Symptom experiences, stress and coping, self-esteem, social support, and HRQoL were measured. Descriptive analysis, the independent t-test, one-way analysis of variance, Pearson correlation analysis, and stepwise multiple regression were used for data analysis. Results: The mean HRQoL score was 72.26 points out of 100. HRQoL showed statistically significant correlations with symptom experiences, stress, self-esteem, social support, satisfaction with peer relationships, and perceived attentiveness of the medical team. The most influential predictors of HRQoL were symptom experiences (β=-.51, p<.001) and satisfaction with peer relationships (β=.32, p<.001). Conclusion: The experiences of different symptoms by children and adolescents who receive HSCT must be considered. In addition, nursing interventions, such as self-help meetings with peer groups, should be provided to improve their HRQoL.