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Yielding and Growing of Adipose Stem Cell Harvested from the Superficial and Deep Abdominal Subcutaneous Fat: A Case 从腹部浅层和深层皮下脂肪中获取的脂肪干细胞的生成和生长:一个案例
Pub Date : 2016-11-26 DOI: 10.4172/2157-7552.1000185
Daegu Son
A 60 year old woman was enrolled in our study. She was 151 cm tall, weighed 73 kg and had high body mass index (33.19 kg/m2). Adipose tissue specimens were taken from her abdomen by surgical excision. ASCs were isolated in primary culture and cell counts were performed by using an automated cell counter (LUNA™). And 5 × 103 ASCs were seeded into DMEM tissue culture flasks and were cultured for 16 days (n=3). Lastly, self-renewal capacity was demonstrated by colony-forming unit fibroblast (CFUF) assays (n=3).
一位60岁的妇女参加了我们的研究。她身高151厘米,体重73公斤,身体质量指数高(33.19公斤/平方米)。手术切除腹部脂肪组织标本。在原代培养中分离ASCs,使用自动细胞计数器(LUNA™)进行细胞计数。将5 × 103株ASCs接种于DMEM组织培养瓶中,培养16 d (n=3)。最后,通过集落形成单位成纤维细胞(CFUF)试验证明了自我更新能力(n=3)。
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引用次数: 1
Immunopathology of Apical Periodontitis and Refractory Cases 根尖牙周炎及难治性病例的免疫病理分析
Pub Date : 2016-11-17 DOI: 10.4172/2157-7552.1000184
E. Matsuzaki, H. Anan, Noriyoshi Matsumoto, J. Hatakeyama, M. Minakami, T. Izumi
Apical periodontitis is a relatively frequently encountered disease in clinical dentistry; however, its pathogenesis and etiology are not easily elucidated. Therefore, it is not always cured, even when carefully following the highest standards of treatment and intractable apical periodontitis may occur. In addition, in long-term root canal treatment of difficult cases with intractable pain, there may be misunderstandings between the dentist and patient. While acute pain is an indispensable symptom in detecting lesions and disease, sustained chronic pain can decrease an individual’s quality of life with various negative outcomes, including decreased motivation to work. Therefore, endodontic treatments and pain control measures for a diseased tooth in intractable apical periodontitis must be developed. This review outlines the progression from the onset of the lesion and examines the immunology of apical periodontitis based on studies of model animals, indicating that interleukin-1β is a key factor in elucidating the disease state and is expected to lead to the development of an effective treatment for refractory cases.
根尖牙周炎是临床上较为常见的疾病。然而,其发病机制和病因尚不清楚。因此,即使遵循最高标准的治疗,也不一定能治愈,难治性根尖牙周炎可能会发生。此外,在长期根管治疗难治性疼痛的病例中,牙医和患者之间可能存在误解。虽然急性疼痛是检测病变和疾病不可或缺的症状,但持续的慢性疼痛会降低个人的生活质量,带来各种负面结果,包括工作动力下降。因此,对难治性根尖牙炎患病牙齿的根管治疗和疼痛控制措施必须发展。本文综述了根尖牙周炎的发病过程,并在模型动物研究的基础上研究了根尖牙周炎的免疫学,表明白细胞介素-1β是阐明疾病状态的关键因素,并有望导致对难治性病例的有效治疗的发展。
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引用次数: 6
Improvement of Gene Expression Studies in the DimethylnitrosamineInduced Liver Fibrosis Model in the Rat Using Selected Reference Genesfor Quantitative Real Time-PCR Analysis 二甲基亚硝胺诱导大鼠肝纤维化模型中基因表达的改进,选择内参基因进行实时定量pcr分析
Pub Date : 2016-11-10 DOI: 10.4172/2157-7552.1000183
D. Rajendran, Gary Phang Siew Siang, Alden Toh Han Hui, K. Chooi
Background: Liver fibrosis is a reaction to chronic liver injury characterized by excessive accumulation of collagen. Due to their importance as biomarkers, the changes in gene expression in the liver during the development of fibrosis and its subsequent outcomes of cirrhosis, neoplasia or resolution are intensely studied. Quantitative realtime PCR (qPCR) with its ability to detect and measure minute amounts of nucleic acids have been increasingly used in these studies. In qPCR, the quantitation of mRNA is relative and the accuracy of results dependent on the reference genes used for standardization. However, many genes studied are normalized against single reference genes, usually housekeeping genes, without adequate justification. Methods: For the dimethylnitrosamine (DMN) induced liver fibrosis rat model, we tested 8 commonly used candidate genes (Actb, Alb, Sdha, B2m, Rn18s, Hprt1, Ppia and Gapdh) to determine their suitability as reference genes. qPCR results were analysed using four commonly used programs; NormFinder, GeNorm, Comparative ΔCt methods and BestKeeper. Result: It was determined that Gapdh and B2m were the most stable genes in normal liver. However, in DMN treated livers, Gapdh and Ppia were the most stably expressed reference genes. We validated these reference genes by using them to normalize the expression of four genes; Tgfb 1, Col1a1, Col3a1 and Tnf known to be highly expressed in liver fibrosis. Conclusion: Gapdh and Ppia are the most suitable reference genes for the normalization of qPCR data in gene expression studies of the liver in the DMN induced liver fibrosis model in the rat. We advise against the use of Actb in this experimental setting because of its low expression stability.
背景:肝纤维化是慢性肝损伤的一种反应,其特征是胶原蛋白的过度积累。由于它们作为生物标志物的重要性,肝脏中基因表达在纤维化发展过程中的变化及其随后的肝硬化、肿瘤或消退的结果被广泛研究。定量实时PCR (qPCR)以其检测和测量微量核酸的能力在这些研究中得到越来越多的应用。在qPCR中,mRNA的定量是相对的,结果的准确性取决于用于标准化的内参基因。然而,许多被研究的基因都是针对单个内参基因(通常是内参基因)进行规范化的,没有足够的理由。方法:对二甲基亚硝胺(DMN)诱导肝纤维化大鼠模型,检测8个常用候选基因(Actb、Alb、Sdha、B2m、Rn18s、Hprt1、Ppia、Gapdh),确定其作为内参基因的适宜性。采用四种常用程序对qPCR结果进行分析;NormFinder, GeNorm, Comparative ΔCt方法和BestKeeper。结果:Gapdh和B2m是正常肝脏中最稳定的基因。然而,在DMN处理的肝脏中,Gapdh和Ppia是最稳定表达的内参基因。我们利用这些内参基因对四个基因的表达进行规范化,从而验证了它们的有效性;已知Tgfb 1, Col1a1, Col3a1和Tnf在肝纤维化中高表达。结论:Gapdh和Ppia是DMN诱导大鼠肝纤维化模型肝脏基因表达研究中qPCR数据归一化最合适的内参基因。我们不建议在这个实验环境中使用Actb,因为它的表达稳定性很低。
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引用次数: 1
Oriented Microstructure in Neural Tissue Engineering: A Review 神经组织工程中的定向微结构研究进展
Pub Date : 2016-11-10 DOI: 10.4172/2157-7552.1000182
F. Ghorbani, A. Zamanian
Regeneration of neural injuries by formation of new axons and myelination to improve quality of patient life is undeniable difficulty in the world in which scientists examined different strategies from ancient times. Recently, 3D tissue engineering scaffolds simulated original extracellular matrix (ECM) and provide desirable substrate for cellular attachment, proliferation and differentiation. However, similarity of scaffold’s materials to ECM contaminant is effective in achieving better results. Investigations demonstrated that oriented fibers, pores and unidirectional channels or conduits act as cell guidance and showed significant effect on cellular differentiation and axonal reconstruction. Between all the methods of scaffold fabrication, freeze casting provides lamellar type and controlled pores that are necessary in neural tissue engineering. In brief, designing scaffolds with oriented structure such as freeze casting with unidirectional solidification and seeding an appropriate cell before implantation improve repair process of neural damages.
通过形成新的轴突和髓鞘来再生神经损伤以提高患者的生活质量是世界上不可否认的困难,科学家们从古代开始就研究了不同的策略。近年来,三维组织工程支架模拟了原始细胞外基质(ECM),为细胞附着、增殖和分化提供了理想的基质。然而,支架材料与ECM污染物的相似性是获得更好效果的有效途径。研究表明,定向纤维、孔隙和单向通道或导管在细胞分化和轴突重建中起着重要的引导作用。在所有的支架制造方法中,冷冻铸造提供了神经组织工程中所必需的层状类型和受控孔隙。简而言之,设计定向结构的支架,如单向凝固的冷冻铸造,在植入前播种合适的细胞,可以改善神经损伤的修复过程。
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引用次数: 8
Human Tissue Engineering: A Relevant Model for Identifying microRNAs Activated by Glioblastoma/Neural Tissue Interaction 人体组织工程:鉴定胶质母细胞瘤/神经组织相互作用激活的microrna的相关模型
Pub Date : 2016-10-29 DOI: 10.4172/2157-7552.1000180
É. Cosset
A key challenge in glioblastoma biology and relevant tumor environment modeling lies in understanding of the cellular interactions occuring within the context of a 3-dimensional (3-D) human tissue microenvironment. Recently, human engineered neural tissue raise the opportunity to study such a very important feature of GBM in vitro. Here, we introduce this model for studying microRNAs.
胶质母细胞瘤生物学和相关肿瘤环境建模的一个关键挑战在于理解在三维(3-D)人体组织微环境中发生的细胞相互作用。最近,人类工程神经组织为在体外研究GBM的这一重要特征提供了机会。在这里,我们介绍了这种研究microrna的模型。
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引用次数: 1
Actovegin equals performance enhancing drug doping: fact or fiction activegin等同于兴奋剂:事实还是虚构
Pub Date : 2016-10-23 DOI: 10.4172/2157-7552.1000179
P. Lee, A. Kwan, Paul M. Smith, James P. Brock, L. Nokes
Editorial article published open access in Journal of Tissue Science and Engineering available at http://dx.doi.org/10.4172/2157-7552.1000179
社论文章发表在组织科学与工程杂志上,可在http://dx.doi.org/10.4172/2157-7552.1000179上获取
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引用次数: 3
Cell culture techniques essential for toxicity testing of inhaled materials and nanomaterials in vitro 细胞培养技术对吸入材料和纳米材料的体外毒性测试至关重要
Pub Date : 2016-10-05 DOI: 10.4172/2157-7552.1000181
S. Bakand
Human tissue is bombarded by a huge range of chemicals. Our lungs are inhaling pollution from both stationary and mobile sources as well as inhaled nanoparticles (NPs) and therapeutic products designed to provide new and innovative medical solutions. Our challenge is to identify what exposures are putting us at risk and balance the risk against benefits that we may receive from these chemicals and new products [1-4]. Advances in in vitro cell culture technology may provide some of the answers.
人体组织受到大量化学物质的轰炸。我们的肺部正在吸入来自固定和移动源的污染,以及吸入的纳米颗粒(NPs)和旨在提供新的创新医疗解决方案的治疗产品。我们面临的挑战是确定哪些暴露使我们面临风险,并平衡风险与我们可能从这些化学品和新产品中获得的益处[1-4]。体外细胞培养技术的进步可能会提供一些答案。
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引用次数: 10
Current Status of Adipose-Derived Mesenchymal Stem Cells in Cartilage Repair 脂肪来源间充质干细胞在软骨修复中的现状
Pub Date : 2016-09-30 DOI: 10.4172/2157-7552.1000177
Arianna L. Gianakos, J. Kennedy
Mesenchymal stem cells (MSCs) have gained increased attention in the treatment of injury-related cartilage damage. Most of the literature has focused on the use of bone marrow MSCs for chondrogensis; however its clinical use has presented consequences of donor site morbidity and pain as well as low cell number during harvest. Adipose tissue offers an abundant source of mesenchymal stem cells (MSCs), showing promise for the treatment of cartilaginous injuries. Numerous studies have focused on the safety and efficacy of adipose-derived mesenchymal stem cells (ADSC) in the form of the stromal vascular fraction (SVF), supporting its use in future clinical applications. While reports of ADSC potential for cartilage repair have been encouraging, the lack of standardization among studies, as well as the confusing terminology describing the different forms of ADSC presents cause for deeper investigation of those studies reporting clinical success. Overall, ADSCs show significant potential as an alternative and/or adjunctive treatment of cartilaginous injuries.
间充质干细胞(MSCs)在损伤相关软骨损伤的治疗中受到越来越多的关注。大多数文献都集中在骨髓间充质干细胞用于软骨形成;然而,它的临床应用已经提出了供体部位的发病率和疼痛的后果,以及低细胞数量在收获。脂肪组织提供了丰富的间充质干细胞(MSCs)来源,显示出治疗软骨损伤的希望。大量研究聚焦于基质血管部分(SVF)形式的脂肪源性间充质干细胞(ADSC)的安全性和有效性,支持其在未来的临床应用。虽然关于ADSC在软骨修复方面的潜力的报道令人鼓舞,但研究之间缺乏标准化,以及描述不同形式ADSC的混淆术语,导致对那些报告临床成功的研究进行更深入的调查。总的来说,ADSCs作为软骨损伤的替代和/或辅助治疗显示出巨大的潜力。
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引用次数: 1
Cell Sheet Technology using Human Umbilical Cord Mesenchymal Stem Cells for Myocardial Tissue Engineering 利用人脐带间充质干细胞进行心肌组织工程的细胞片技术
Pub Date : 2016-09-11 DOI: 10.4172/2157-7552.1000178
Lakshmi R Nair, T. Kumary
With limited regenerative capacity and most complex structural and electrophysiological properties; recapitulating the cardiac tissue is a challenging task for the researchers. The cell injection was found unreliable due to the cell loss and low retention of the transplanted cells. This could be overcome by the technique of cell sheet engineering. Scaffold free, thick, cell dense, three dimensional constructs could be generated for suture free transplantation. For the generation of the cardiac constructs the neonatal cardiomyocytes and myoblasts were mostly used. They were tedious to isolate and culture and the risk of arrhythmogenic foci prevailed. Hence the concept of differentiating a suitable allogeneic cell source to myocardial lineage seemed relevant. Human umbilical cord mesenchymal stem cells (hUCMSCs) are emerging with the assistance of differentiating agents and cell sheet engineering for addressing the cardiac regeneration. The preliminary report on this regard has been published. The cells attained cardiomyocyte-like morphology with the expression of alpha-actinin and myosin heavy chain on culturing with cardiac conditioned medium and the inducer sphingosine-1-phosphate. It presented cardiomyocyte-like action potential and voltage gated currents. Hence the cell sheet engineering approach with cells differentiated to cardiac lineage using specific agents is a recent area to be explored.
具有有限的再生能力和最复杂的结构和电生理特性;对研究人员来说,再现心脏组织是一项具有挑战性的任务。由于细胞丢失和移植细胞的低保留率,细胞注射被发现是不可靠的。这可以通过细胞片工程技术来克服。无支架、粗大、细胞致密、三维结构可用于无缝线移植。在心脏构建体的生成中,主要采用新生儿心肌细胞和成肌细胞。分离培养过程繁琐,易发生心律失常灶。因此,将合适的同种异体细胞来源分化为心肌谱系的概念似乎是相关的。人脐带间充质干细胞(hUCMSCs)是在分化剂和细胞片工程的帮助下出现的,用于心脏再生。关于这方面的初步报告已经发表。在心脏条件培养基和诱导剂鞘氨醇-1-磷酸的作用下,细胞表达α -肌动蛋白和肌球蛋白重链,形成心肌细胞样形态。表现为心肌细胞样动作电位和电压门控电流。因此,利用特定试剂将细胞分化为心脏谱系的细胞片工程方法是一个有待探索的新领域。
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引用次数: 3
Efficient Plant Regeneration of Selected Kenyan Sweetpotato (Ipomoea batatas (L.) Lam.) Cultivars through Somatic Embryogenesis 肯尼亚甘薯的高效植株再生研究Lam)。通过体细胞胚胎发生的品种
Pub Date : 2016-08-25 DOI: 10.4172/2157-7552.1000176
W. Mbinda, S. Anami, O. Ombori, C. Dixelius, Richard O. Oduor
Sweetpotato is an important food crop in the world as well as in Kenya. Various fungal, and viral diseases are major constraints in its production and are currently threatening the sweetpotato production in sub-Saharan Africa. Genetic engineering offers significant potential for the crop’s genetic improvement. However, this is limited by the low efi¬Âciency and strong genotype dependency in tissue culture. This study aimed to establish an efficient somatic embryogenesis and plant regeneration system using shoot apical meristem explants of sweetpotato. Three sweetpotato cultivars that are widely grown in Kenya; KSP36, Kemb36 and Mweu mutheke along with an exotic model cultivar Jewel were evaluated. The maximum somatic embryogenic induction, at 96.72%, was obtained from explants cultured on Linsmaier and Skoog salts and vitamins medium supplemented with 0.5 mg/l dichlorophenoxyacetic acid and 0.2 mg/l zeatin riboside. The highest number of shoot induction (33) was observed after transfer of embryonic callus to embryo maturation medium supplemented with 2 mg/l abscisic acid. Significant differences were observed between cultivars for somatic embryogenesis and plant regeneration. Jewel showed the best response, while Mweu mutheke was the least responsive under the culture conditions tested in this study. Regenerated plants were successfully rooted and grown to maturity after hardening in soil in the greenhouse. Such a robust, successful and efi¬Âcient system possesses the potential to become an important tool for crop improvement and functional studies of genes in sweetpotato.
甘薯是世界和肯尼亚重要的粮食作物。各种真菌和病毒疾病是其生产的主要制约因素,目前正威胁着撒哈拉以南非洲的甘薯生产。基因工程为作物的遗传改良提供了巨大的潜力。然而,这受到组织培养效率低、Â效率高和基因型依赖性强的限制。本研究旨在利用甘薯茎尖分生组织外植体建立高效的体细胞胚胎发生和植株再生体系。肯尼亚广泛种植的三种甘薯品种;以KSP36、Kemb36和Mweu mutheke以及一个外来模式品种Jewel为研究对象。在添加0.5 mg/l二氯苯氧乙酸和0.2 mg/l玉米蛋白核苷的Linsmaier盐和Skoog盐和维生素培养基上培养的外植体胚诱导率最高,为96.72%。将胚愈伤组织转移到添加2 mg/l脱落酸的胚成熟培养基中,诱导芽数最多(33个)。在体细胞胚发生和植株再生方面,不同品种间存在显著差异。在本研究测试的培养条件下,Jewel的反应最好,而Mweu mutheke的反应最差。再生植株在温室土壤硬化后成功生根并生长成熟。这样一个强大、成功和高效的系统有潜力成为甘薯作物改良和基因功能研究的重要工具。
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引用次数: 8
期刊
Journal of Tissue Science and Engineering
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