• Title/Summary/Keyword: Cell fate

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Effects of Extracellular Matrix Protein-derived Signaling on the Maintenance of the Undifferentiated State of Spermatogonial Stem Cells from Porcine Neonatal Testis

  • Park, Min Hee;Park, Ji Eun;Kim, Min Seong;Lee, Kwon Young;Hwang, Jae Yeon;Yun, Jung Im;Choi, Jung Hoon;Lee, Eunsong;Lee, Seung Tae
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.10
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    • pp.1398-1406
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    • 2016
  • In general, the seminiferous tubule basement membrane (STBM), comprising laminin, collagen IV, perlecan, and entactin, plays an important role in self-renewal and spermatogenesis of spermatogonial stem cells (SSCs) in the testis. However, among the diverse extracellular matrix (ECM) proteins constituting the STBM, the mechanism by which each regulates SSC fate has yet to be revealed. Accordingly, we investigated the effects of various ECM proteins on the maintenance of the undifferentiated state of SSCs in pigs. First, an extracellular signaling-free culture system was optimized, and alkaline phosphatase (AP) activity and transcriptional regulation of SSC-specific genes were analyzed in porcine SSCs (pSSCs) cultured for 1, 3, and 5 days on non-, laminin- and collagen IV-coated Petri dishes in the optimized culture system. The microenvironment consisting of glial cell-derived neurotrophic factor (GDNF)-supplemented mouse embryonic stem cell culture medium (mESCCM) (GDNF-mESCCM) demonstrated the highest efficiency in the maintenance of AP activity. Moreover, under the established extracellular signaling-free microenvironment, effective maintenance of AP activity and SSC-specific gene expression was detected in pSSCs experiencing laminin-derived signaling. From these results, we believe that laminin can serve as an extracellular niche factor required for the in vitro maintenance of undifferentiated pSSCs in the establishment of the pSSC culture system.

HSP27 EXPRESSION IN OSTEOBLAST BY THERMAL STRESS (골모세포에서 열자극에 의한 Hsp27 발현에 대한 연구)

  • Rim, Jae-Suk;Kim, Byeong-Ryol;Kwon, Jong-Jin;Jang, Hyon-Seok;Lee, Eui-Suk;Jun, Sang-Ho;Woo, Hyeon-Il
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.30 no.1
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    • pp.11-21
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    • 2008
  • Aim of the study: Thermal stress is a central determinant of osseous surgical outcomes. Interestingly, the temperatures measured during endosseous surgeries coincide with the temperatures that elicit the heat shock response of mammalian cells. The heat shock response is a coordinated biochemical response that helps to protect cells from stresses of various forms. Several protective proteins, termed heat shock proteins (hsp) are produced as part of this response. To begin to understand the role of the stress response of osteoblasts during surgical manipulation of bone, the heat shock protein response was evaluated in osteoblastic cells. Materials & methods: With primary cell culture studies and ROS 17/2.8 osteoblastic cells transfected with hsp27 encoding vectors culture studies, the thermal stress response of mammalian osteoblastic cells was evaluated by immunohistochemistry and western blot analysis. Results: Immunocytochemistry indicated that hsp27 was present in unstressed osteoblastic cells, but not fibroblastic cells. Primarily cultured osteoblasts and fibroblasts expressed the major hsp in response to thermal stress, however, the small Mr hsp, hsp27 was shown to be a constitutive product only in osteoblasts. Creation of stable transformed osteoblastic cells expressing abundant hsp27 protein was used to demonstrate that hsp27 confers stress resistance to osteoblastic cells. Conclusions: The demonstrable presence and function of hsp27 in cultured bones and cells implicates this protein as a determinant of osteoblastic cell fate in vivo.

Decreases in $Casz1$ mRNA by an siRNA Complex Do not Alter Blood Pressure in Mice

  • Ji, Su-Min;Shin, Young-Bin;Park, So-Yon;Lee, Hyeon-Ju;Oh, Berm-Seok
    • Genomics & Informatics
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    • v.10 no.1
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    • pp.40-43
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    • 2012
  • Recent genomewide association studies of large samples have identified genes that are associated with blood pressure. The Global Blood Pressure Genetics (Global BPgen) and Cohorts for Heart and Aging Research in Genome Epidemiology (CHARGE) consortiums identified 14 loci that govern blood pressure on a genomewide significance level, one of which is $CASZ1$ confirmed in both Europeans and Asians. $CASZ1$ is a zinc finger transcription factor that controls apoptosis and cell fate and suppresses neuroblastoma tumor growth by reprogramming gene expression, like a tumor suppressor. To validate the function of $CASZ1$ in blood pressure, we decreased $Casz1$ mRNA levels in mice by siRNA. $Casz1$ siRNA reduced mRNA levels by 59% in a mouse cell line. A polyethylenimine-mixed siRNA complex was injected into mouse tail veins, reducing $Casz1$ mRNA expression to 45% in the kidney. However, blood pressure in the treated mice was unaffected, despite a 55% reduction in $Casz1$ mRNA levels in the kidney on multiple siRNA injections daily. Even though $Casz1$ siRNA-treated mice did not experience any significant change in blood pressure, our study demonstrates the value of $in$ $vivo$ siRNA injection in analyzing the function of candidate genes identified by genomewide association studies.

Infection Mechanism of Pathogenic Exduate by Soil-Borne Fungal Pathogens : A Review

  • Lim, You-Jin;Kim, Hye-Jin;Song, Jin-A;Chung, Doug-Young
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.4
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    • pp.622-627
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    • 2012
  • The processes to determine the composition, dynamics, and activity of infection mechanisms by the rhizosphere microflora have attracted the interest of scientists from multiple disciplines although considerable progress of the infection pathways and plant-pathogen interactions by soil borne fungal pathogens have been made. Soilborne pathogens are confined within a three-dimensional matrix of mineral soil particles, pores, organic matter in various stages of decomposition and a biological component. Among the physical and chemical properties of soils soil texture and matric water potential may be the two most important factors that determine spread exudates by soil borne fungal pathogens, based on the size of the soil pores. Pathogenic invasion of plant roots involves complex molecular mechanisms which occur in the diffuse interface between the root and the soil created by root exudates. The initial infection by soilborne pathogens can be caused by enzymes which breakdown cell wall layers to penetrate the plant cell wall for the fungus. However, the fate and mobility of the exudates are less well understood. Therefore, it needs to develop methods to control disease caused by enzymes produced by the soilborne pathogens by verifying many other possible pathways and mechanisms of infection processes occurring in soils.

Subretinal transplantation of putative retinal pigment epithelial cells derived from human embryonic stem cells in rat retinal degeneration model

  • Park, Un-Chul;Cho, Myung-Soo;Park, Jung-Hyun;Kim, Sang-Jin;Ku, Seung-Yup;Choi, Young-Min;Moon, Shin-Yong;Yu, Hyeong-Gon
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.4
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    • pp.216-221
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    • 2011
  • Objective: To differentiate the human embryonic stem cells (hESCs) into the retinal pigment epithelium (RPE) in the defined culture condition and determine its therapeutic potential for the treatment of retinal degenerative diseases. Methods: The embryoid bodies were formed from hESCs and attached on the matrigel coated culture dishes. The neural structures consisting neural precursors were selected and expanded to form rosette structures. The mechanically isolated neural rosettes were differentiated into pigmented cells in the media comprised of N2 and B27. Expression profiles of markers related to RPE development were analyzed by reverse transcription-polymerase chain reaction and immunostaining. Dissociated putative RPE cells ($10^5$ cells/5 ${\mu}L$) were transplanted into the subretinal space of rat retinal degeneration model induced by intravenous sodium iodate injection. Animals were sacrificed at 1, 2, and 4 weeks after transplantation, and immnohistochemistry study was performed to verify the survival of the transplanted cells. Results: The putative RPE cells derived from hESC showed characteristics of the human RPE cells morphologically and expressed molecular markers and associated with RPE fate. Grafted RPE cells were found to survive in the subretinal space up to 4 weeks after transplantation, and the expression of RPE markers was confirmed with immunohistochemistry. Conclusion: Transplanted RPE cells derived from hESC in the defined culture condition successfully survived and migrated within subretinal space of rat retinal degeneration model. These results support the feasibility of the hESC derived RPE cells for cell-based therapies for retinal degenerative disease.

Effects of 915 MHz Radiofrequency Identification Electromagnetic Field Exposure on Neuronal Precursor Cells in the Dentate Gyrus of Adult Rat Brains

  • Kim, Hye Sun;Lee, Yu Hee;Lee, Yun-Sil;Choi, Hyung-Do;Kwon, Jong Hwa;Pack, Jeong-Ki;Kim, Nam;Ahn, Young Hwan
    • Journal of electromagnetic engineering and science
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    • v.15 no.3
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    • pp.173-180
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    • 2015
  • To explore the effects of radiofrequency electromagnetic field on the fate of neuronal cells, we investigated whether exposure to 915 MHz radiofrequency identification (RFID) caused morphological changes in neuronal cells in rat hippocampal dentate gyrus (DG). A reverberation chamber was used as a whole-body RFID exposure system. Rats were assigned to two groups: sham- and RFID-exposed groups. Rats in the RFID-exposed group were exposed to RFID at 4 W/kg specific absorption rate (SAR) for 8 hours daily, 5 days per week, for 2 weeks. Morphological evaluation of DG was performed using immunohistochemistry with doublecortin (DCX) as a neuronal precursor cell marker and neuronal nuclei (NeuN) as a mature neuronal cell marker. No significant morphological changes in DCX+ or NeuN+ cells in the DG of RFID-exposed rats were observed. These results suggest that RFID exposure induces no significant change in DCX+ neuronal precursor or NeuN+ neuronal cells in DG of rats.

High-Speed Reed-Solomon Decoder Using New Degree Computationless Modified Euclid´s Algorithm (새로운 DCME 알고리즘을 사용한 고속 Reed-Solomon 복호기)

  • 백재현;선우명훈
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.40 no.6
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    • pp.459-468
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    • 2003
  • This paper proposes a novel low-cost and high-speed Reed-Solomon (RS) decoder based on a new degree computationless modified Euclid´s (DCME) algorithm. This architecture has quite low hardware complexity compared with conventional modified Euclid´s (ME) architectures, since it can remove completely the degree computation and comparison circuits. The architecture employing a systolic away requires only the latency of 2t clock cycles to solve the key equation without initial latency. In addition, the DCME architecture using 3t+2 basic cells has regularity and scalability since it uses only one processing element. The RS decoder has been synthesized using the 0.25${\mu}{\textrm}{m}$. Faraday CMOS standard cell library and operates at 200MHz and its data rate suppots up to 1.6Gbps. For tile (255, 239, 8) RS code, the gate counts of the DCME architecture and the whole RS decoder excluding FIFO memory are only 21,760 and 42,213, respectively. The proposed RS decoder can reduce the total fate count at least 23% and the total latency at least 10% compared with conventional ME architectures.

Apple pectin, a dietary fiber, ameliorates myocardial injury by inhibiting apoptosis in a rat model of ischemia/reperfusion

  • Lim, Sun Ha;Kim, Mi Young;Lee, Jongwon
    • Nutrition Research and Practice
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    • v.8 no.4
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    • pp.391-397
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    • 2014
  • BACKGROUND/OBJECTIVE: Myocardial cell death due to occlusion of the coronary arteries leads to myocardial infarction, a subset of coronary heart disease (CHD). Dietary fiber is known to be associated with a reduced risk of CHD, the underlying mechanisms of which were suggested to delay the onset of occlusion by ameliorating risk factors. In this study, we tested a hypothesis that a beneficial role of dietary fiber could arise from protection of myocardial cells against ischemic injury, manifested after occlusion of the arteries. MATERIALS/METHODS: Three days after rats were fed apple pectin (AP) (with 10, 40, 100, and 400 mg/kg/day), myocardial ischemic injury was induced by 30 min-ligation of the left anterior descending coronary artery, followed by 3 hr-reperfusion. The area at risk and infarct area were evaluated using Evans blue dye and 2,3,5-triphenyltetrazolium chloride (TTC) staining, respectively. DNA nicks reflecting the extent of myocardial apoptosis were assessed by TUNEL assay. Levels of cleaved caspase-3, Bcl-2, and Bax were assessed by immunohistochemistry. RESULTS: Supplementation of AP (with 100 and 400 mg/kg/day) resulted in significantly attenuated infarct size (IS) (ratio of infarct area to area at risk) by 21.9 and 22.4%, respectively, in the AP-treated group, compared with that in the control group. This attenuation in IS showed correlation with improvement in biomarkers involved in the apoptotic cascades: reduction of apoptotic cells, inhibition of conversion of procaspase-3 to caspase-3, and increase of Bcl-2/Bax ratio, a determinant of cell fate. CONCLUSIONS: The findings indicate that supplementation of AP results in amelioration of myocardial infarction by inhibition of apoptosis. Thus, the current study suggests that intake of dietary fiber reduces the risk of CHD, not only by blocking steps leading to occlusion, but also by protecting against ischemic injury caused by occlusion of the arteries.

Micropatterning on Biodegradable Nanofiber Scaffolds by Femtosecond Laser Ablation Process (펨토초 레이저 절삭 공정을 이용한 생분해성 나노섬유 표면 미세 패터닝 공정)

  • Chung, Yongwoo;Jun, Indong;Kim, Yu-Chan;Seok, Hyun-Kwang;Chung, Seok;Jeon, Hojeong
    • Journal of the Korean institute of surface engineering
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    • v.49 no.6
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    • pp.555-559
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    • 2016
  • A biodegradable nanofiber scaffolds using electrospining provide fibrous guidance cues for controlling cell fate that mimic the native extracellular matrix (ECM). It can create a pattern using conventional electrospining method, but has a difficulty to generate one or more pattern structures. Femtosecond(fs) laser ablation has much interested in patterning on biomaterials in order to distinguish the fundamental or systemic interaction between cell and material surface. The ablated materials with a short pulse duration using femtosecond laser that allows for precise removal of materials without transition of the inherent material properties. In this study, linear grooves and circular craters were fabricated on electrospun nanofiber scaffolds (poly-L-lactide(PLLA)) by femtosecond laser patterning processes. As parametric studies, pulse energy and beam spot size were varied to determine the effects of the laser pulse on groove size. We confirmed controlling pulse energy to $5{\mu}J-20{\mu}J$ and variation of lens maginfication of 2X, 5X, 10X, 20X created grooves of width to approximately $5{\mu}m-50{\mu}m$. Our results demonstrate that femtosecond laser processing is an effective means for flexibly structuring the surface of electrospun PLLA nanofibers.

The CCAAT-box transcription factor, NF-Y complex, mediates the specification of the IL1 neurons in C. elegans

  • Woojung Heo;Hyeonjeong Hwang;Jimin Kim;Seung Hee Oh;Youngseok Yu;Jae-Hyung Lee;Kyuhyung Kim
    • BMB Reports
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    • v.56 no.3
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    • pp.153-159
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    • 2023
  • Neuronal differentiation is highly coordinated through a cascade of gene expression, mediated via interactions between trans-acting transcription factors and cis-regulatory elements of their target genes. However, the mechanisms of transcriptional regulation that determine neuronal cell-fate are not fully understood. Here, we show that the nuclear transcription factor Y (NF-Y) subunit, NFYA-1, is necessary and sufficient to express the flp-3 neuropeptide gene in the IL1 neurons of C. elegans. flp-3 expression is decreased in dorsal and lateral, but not ventral IL1s of nfya-1 mutants. The expression of another terminally differentiated gene, eat-4 vesicular glutamate transporter, is abolished, whereas the unc-8 DEG/ENaC gene and pan-neuronal genes are expressed normally in IL1s of nfya-1 mutants. nfya-1 is expressed in and acts in IL1s to regulate flp-3 and eat-4 expression. Ectopic expression of NFYA-1 drives the expression of flp-3 gene in other cell-types. Promoter analysis of IL1-expressed genes results in the identification of several cis-regulatory motifs which are necessary for IL1 expression, including a putative CCAAT-box located in the flp-3 promoter that NFYA-1 directly interacts with. NFYA-1 and NFYA-2, together with NFYB-1 and NFYC-1, exhibit partly or fully redundant roles in the regulation of flp-3 or unc-8 expression, respectively. Taken together, our data indicate that the NF-Y complex regulates neuronal subtype-specification via regulating a set of terminal-differentiation genes.