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Preservation of Microbial Pure Cultures and Mixed Communities 微生物纯培养物和混合群落的保存
Pub Date : 2015-07-31 DOI: 10.1007/8623_2015_51
Bram Vekeman, K. Heylen
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引用次数: 11
The outcome of Minibiobank under adverse conditions in Iraq 微型生物银行在伊拉克不利条件下的结果
Pub Date : 2015-07-31 DOI: 10.4172/2157-7552.S1.019
Mohemid Maddallah Mohammed AlJebouri
T University of Arizona Biorepository is responsible for collecting, storing, tracking, processing, and distributing human tissue, blood, and other bio-specimens. The Biorepository’s mission is to provide high-quality, clinically annotated specimens to the research community at this and other institutions as well as to industry involved in biomedical research. Researchers can use the stored materials for future research studies to learn more about cancer, diabetes, and other health problems. The bank will provide a ready supply of samples, so researchers do not have to look for donors for each new study. The biorepository provides consistent collection, processing, banking and clinical correlative procedures and operates in conjunction with departments of Surgery, Ophthalmology, Pulmonology, Aging, Neurology, Neurosurgery, Biomedical Engineering, OB-GYN, Cardiology and others. Currently the bio-repository has in its possession 1.5 million patient samples obtained through Pathology. The bio-repository utilizes universal electronic consents providing annotated clinical data on each patient through an Honest Broker arrangement. All samples are linked to patient medical identifiers allowing for access to the electronic health record now and at all times in the future in a deidentified fashion. Fresh frozen paraffin blocks, blood, plasma, sera, urine and other biological fluids; as well as biopsies and other tissues/bio-samples are stored. In addition, DNA/RNA/proteins are also banked for all samples. Significantly, we have now developed methodology allowing for genomic and phenotypic sample characterization for approx. $2/patient sample. As all samples are stored according to industry best practices, in temperature-controlled, monitored, and alarmed environments (in LN2 or other freezers as appropriate) to maximize resource integrity utilizing cGTP practices whenever possible, the biorepository also serves as a source of cells and tissues for projects involving translational and regenerative medicine. To facilitate investigator interactions the biorepository utilizes Tissue Metrix2 as the central database for information on all banked bio-samples. The software has a web-based front-end with an Oracle database which permits access from web browsers across multiple platforms. It employs role based security to permit control over user access to information stored in the database. The i2b2 open source exploration tool is used as a storefront for investigator sample requests in a text based format.
亚利桑那大学生物储存库负责收集、储存、跟踪、处理和分发人体组织、血液和其他生物标本。生物储存库的使命是为本院和其他机构的研究界以及涉及生物医学研究的行业提供高质量的临床注释标本。研究人员可以将储存的材料用于未来的研究,以了解更多关于癌症、糖尿病和其他健康问题的信息。该银行将提供现成的样本供应,因此研究人员不必为每项新研究寻找捐赠者。生物储存库提供一致的收集、处理、存储和临床相关程序,并与外科、眼科、肺科、老龄化、神经病学、神经外科、生物医学工程、妇产科、心脏病学等部门合作。目前,生物信息库拥有通过病理学获得的150万例患者样本。生物资源库利用通用电子同意,通过诚实经纪人安排为每位患者提供注释的临床数据。所有样本都与患者医疗标识符相关联,允许现在和将来任何时候以不识别的方式访问电子健康记录。新鲜冷冻石蜡块、血液、血浆、血清、尿液等生物体液;以及活组织检查和其他组织/生物样本被储存。此外,所有样品的DNA/RNA/蛋白质也被储存起来。值得注意的是,我们现在已经开发了一种方法,允许基因组和表型样品表征大约。2美元/患者样本。由于所有样品都按照行业最佳实践保存,在温度控制、监测和警报环境中(在LN2或其他适当的冷冻室中),尽可能利用cGTP实践最大限度地提高资源完整性,生物库还可作为涉及转化和再生医学项目的细胞和组织的来源。为了方便研究者的互动,生物库利用组织Metrix2作为所有生物样本信息的中心数据库。该软件有一个基于web的前端,使用Oracle数据库,允许跨多个平台的web浏览器访问。它采用基于角色的安全性来控制用户对存储在数据库中的信息的访问。i2b2开源探索工具被用作基于文本格式的调查员样本请求的店面。
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引用次数: 5
Role of immunohistochemistry and biobanking in targeted cancer therapy 免疫组织化学和生物银行在肿瘤靶向治疗中的作用
Pub Date : 2015-07-31 DOI: 10.4172/2157-7552.S1.018
B. Kumar
C which has been the standard therapeutic regimen for cancer has the disadvantages of conforming to the “one size fits all” style. These standard drugs fail to distinguish malignant versus normal tissue, thus bringing along a range of adverse effects. Targeted treatment on the contrary show a greater selectivity for tumor cells and causes less damage to normal cells. It is to be noted that morphologically distinct tumours show variable biological characteristics and response to treatment. It is thus becoming important to identify these targets within the cancer tissue which include the tumour cells and the tumour microenvironment (ie, stromal cells, microvessels, and host’s immune cells), all of which could serve as potential treatment targets.
C一直是癌症的标准治疗方案,但它的缺点是符合“一刀切”的风格。这些标准药物无法区分恶性组织和正常组织,从而带来一系列不良反应。相反,靶向治疗对肿瘤细胞有更大的选择性,对正常细胞的损伤更小。值得注意的是,形态不同的肿瘤表现出不同的生物学特征和对治疗的反应。因此,在肿瘤组织内识别这些靶点(包括肿瘤细胞和肿瘤微环境(即基质细胞、微血管和宿主免疫细胞))变得非常重要,所有这些靶点都可以作为潜在的治疗靶点。
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引用次数: 0
Development of Novel, Bioresorbable, Small-Diameter Electrospun Vascular Grafts 新型、生物可吸收、小直径电纺丝血管移植物的发展
Pub Date : 2015-03-25 DOI: 10.4172/2157-7552.1000151
Jed Johnson, Devan Ohst, Tyler Groehl, Sarah Hetterscheidt, Matthew Jones
This study proposes a production method capable of producing vascular grafts from fully synthetic, resorbable polymers that both meet basic minimum mechanical requirements for potential vascular grafts, and have a compliance similar to that of the intended vasculature being replaced. All of the electrospun vascular grafts in this work meet the minimum mechanical requirements for compliance, burst pressure, and suture retention strength, and could be potential candidates for off-the-shelf tissue engineered vascular grafts. Each polymer investigated in this paper has FDA approval for medical use and has been shown to be successful in various tissue engineering applications. Only recently has an electrospun small-diameter graft been fabricated with compliance and burst pressure greater than that of the human saphenous vein. We show a significant advancement in burst pressure, compliance, and suture retention strength in the novel electrospun grafts presented in this work which demonstrates the potential use of these tissue engineered vascular grafts for coronary artery bypass graft and other smalldiameter graft indications.
本研究提出了一种生产方法,能够用完全合成的、可吸收的聚合物生产血管移植物,这种聚合物既满足潜在血管移植物的基本最低机械要求,又具有与拟替换的血管相似的顺应性。在这项工作中,所有的电纺丝血管移植物都满足顺应性、破裂压力和缝合保持强度的最低机械要求,可能是现成的组织工程血管移植物的潜在候选者。本文研究的每种聚合物都已获得FDA批准用于医疗用途,并已在各种组织工程应用中取得成功。直到最近才有一种小直径的电纺丝移植物被制造出来,其顺应性和破裂压力大于人类隐静脉。我们展示了在破裂压力、顺应性和缝线保持强度方面的重大进步,在这项工作中提出的新型电纺丝血管移植物,证明了这些组织工程血管移植物在冠状动脉旁路移植和其他小直径移植物适应症中的潜在应用。
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引用次数: 25
Cellular Approaches to Tissue-Engineering of Skin: A Review 皮肤组织工程的细胞方法:综述
Pub Date : 2015-03-23 DOI: 10.4172/2157-7552.1000150
Wei Long Ng, W. Yeong, M. Naing
The human skin is a complex organ consisting of multiple skin cells that work together to complement each other and provide essential functions such as skin barrier function, skin homeostasis and protection against the harmful ultraviolet radiation. Understanding the roles and paracrine signaling of different skin cells plus the influence of external stimuli on them are crucial towards the design of tissue-engineered skin constructs as these factors regulate the cellular behavior such as cell proliferation, migration and differentiation. Hence, an in-depth understanding of the knowledge on the epithelial-mesenchymal interactions would be valuable towards the design of a tissue-engineered skin construct.
人体皮肤是一个复杂的器官,由多个皮肤细胞组成,这些细胞协同工作,相互补充,并提供皮肤屏障功能,皮肤稳态和防止有害紫外线辐射等基本功能。了解不同皮肤细胞的作用和旁分泌信号以及外部刺激对它们的影响对组织工程皮肤结构的设计至关重要,因为这些因素调节细胞行为,如细胞增殖、迁移和分化。因此,深入了解上皮-间充质相互作用的知识对组织工程皮肤结构的设计是有价值的。
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引用次数: 18
Decellularization of Human Lip 人嘴唇的脱细胞
Pub Date : 2015-02-26 DOI: 10.4172/2157-7552.1000149
H. Khan, M. Szarko, P. Butler
Introduction: Perioral tissue loss commonly requires surgical reconstruction. Autologous tissue transfer result in poor functional and aesthetic outcomes and allogenous transplantation of tissue requires lifelong immunosuppression. There is a clinical need for a cell scaffold, which could be seeded with the patients own cells to create an immunogenically inert perioral tissue replacement. Decellularized human lip may provide the answer. Methodology: To the authors knowledge this was the first time human lip has been decellularized. Four existing protocols shown to be successful at removing cells from either muscle or dermis were used to decellularize human lip in an attempt to identify an optimal protocol. Results: Three of the four protocols proved to be successful at achieving decellularization of the lip, as histological investigation of these samples showed complete loss of cellular structures for the entire construct. A non-detergent based protocol using osmotic shock and enzymatic processes best preserved the extracellular matrix. It was able to maintain the micro-architecture of collagen and elastin, and retain important signaling molecules such as glycosaminoglycans. Conclusion: This decellularized scaffold developed here may be the first step towards an exciting new treatment for perioral tissue loss.
口腔周围组织丢失通常需要手术重建。自体组织移植导致较差的功能和美观效果,异体组织移植需要终身免疫抑制。临床需要一种细胞支架,它可以用患者自己的细胞播种,以产生免疫原性惰性的口周组织替代物。去细胞化的人类嘴唇可能提供了答案。方法:据作者所知,这是第一次人类嘴唇被脱细胞。现有的四种方法被证明可以成功地从肌肉或真皮中移除细胞,并被用于人类嘴唇的脱细胞,以试图确定最佳的方法。结果:四种方案中的三种被证明是成功地实现了唇部的脱细胞,因为这些样品的组织学调查显示整个结构的细胞结构完全丧失。使用渗透冲击和酶促过程的非洗涤剂为基础的方案最好地保存了细胞外基质。它能够维持胶原蛋白和弹性蛋白的微观结构,并保留重要的信号分子,如糖胺聚糖。结论:这种脱细胞支架可能是一种令人兴奋的治疗口腔周围组织丢失的新方法的第一步。
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引用次数: 0
Development, Characterization and Cell Cultural Response of 3D Biocompatible Micro-Patterned Poly-ε- Caprolactone Scaffolds Designed and Fabricated Integrating Lithography and Micromolding Fabrication Techniques. 集成光刻和微成型技术设计和制造的三维生物相容性微图案聚ε-己内酯支架的开发、表征和细胞培养反应。
Pub Date : 2015-01-01 DOI: 10.4172/2157-7552.1000145
T. Limongi, E. Miele, V. Shalabaeva, R. Rocca, Rossana Schipani, N. Malara, F. Angelis, A. Giugni, E. Fabrizio
Scaffold design and fabrication are very important subjects for biomaterial, tissue engineering and regenerative medicine research playing a unique role in tissue regeneration and repair. Among synthetic biomaterials Poly-e- Caprolactone (PCL) is very attractive bioresorbable polyester due to its high permeability, biodegradability and capacity to be blended with other biopolymers. Thanks to its ability to naturally degrade in tissues, PCL has a great potential as a new material for implantable biomedical micro devices. This work focuses on the establishment of a micro fabrication process, by integrating lithography and micromolding fabrication techniques, for the realization of 3D microstructure PCL devices. Scaffold surface exhibits a combination in the patterned length scale; cylindrical pillars of 10 μm height and 10 μm diameter are arranged in a hexagonal lattice with periodicity of 30 μm and their sidewalls are nano-sculptured, with a regular pattern of grooves leading to a spatial modulation in the z direction. In order to demonstrate that these biocompatible pillared PCL substrates are suitable for a proper cell growth, NIH/3T3 mouse embryonic fibroblasts were seeded on them and cells key adhesion parameters were evaluated. Scanning Electron Microscopy and immunofluorescence analysis were carried out to check cell survival, proliferation and adhesion; cells growing on the PCL substrates appeared healthy and formed a well-developed network in close contact with the micro and nano features of the pillared surface. Those 3D scaffolds could be a promising solution for a wide range of applications within tissue engineering and regenerative medicine applications.
支架的设计与制造是生物材料、组织工程和再生医学研究的重要课题,在组织再生和修复中发挥着独特的作用。聚己内酯(PCL)具有高渗透性、可生物降解性和与其他生物聚合物共混的能力,是一种非常有吸引力的生物可吸收聚酯材料。由于其在组织中自然降解的能力,PCL作为植入式生物医学微设备的新材料具有很大的潜力。本工作的重点是建立一种微制造工艺,通过集成光刻和微成型制造技术,实现三维微结构的PCL器件。支架表面在图案长度尺度上呈现组合;高10 μm、直径10 μm的圆柱柱呈周期性30 μm的六角形晶格排列,其侧壁进行纳米雕刻,具有规则的凹槽图案,导致z方向的空间调制。为了证明这些生物相容性柱状PCL基质适合细胞生长,我们将NIH/3T3小鼠胚胎成纤维细胞接种在其上,并对细胞的关键粘附参数进行了评估。扫描电镜和免疫荧光检测细胞存活、增殖和粘附情况;在PCL基质上生长的细胞表现健康,并与柱状表面的微纳特征密切接触,形成了发育良好的网络。这些3D支架在组织工程和再生医学的广泛应用中是一个很有前途的解决方案。
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引用次数: 5
Effect of Different Solvents in Solvent Casting of Porous caffolds in Biomedical and Tissue Engineering Applications 不同溶剂对生物医学和组织工程中多孔支架溶剂铸造的影响
Pub Date : 2015-01-01 DOI: 10.4172/2157-7552.1000142
Mousam Choudhury, S. Mohanty, S. Nayak
In the present investigation, PLA porous scaffolds was fabricated using NaCl as porogen in three different solvents i.e. 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP), dichloromethane (DCM) and chloroform (CF) respectively, by solvent casting particulate leaching method. The morphology, structure and thermal behaviour of the PLA scaffolds for porosity measurement were evaluated using scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR); thermo gravimetric analysis (TGA) and differential scanning calorimetry (DSC). The developed porous scaffolds were further characterized for porosity measurement, solvent uptake property and water absorption capacity at different temperatures. PLA/ CF scaffold depicted higher porosity factor (93%) along with enhanced water uptake capacity (220%) as against PLA/ HFIP scaffold (75%). However PLA/ DCM scaffolds illustrated more thermal stability as compared with PLA/ HFIP and PLA/ CF scaffolds.
采用溶剂铸造颗粒浸出法,在1,1,1,3,3,3-六氟-2-丙醇(HFIP)、二氯甲烷(DCM)和氯仿(CF)三种不同溶剂中,以NaCl为孔隙剂制备了聚乳酸多孔支架。采用扫描电镜(SEM)、傅里叶变换红外光谱(FTIR)对PLA支架的形貌、结构和热行为进行了表征;热重分析(TGA)和差示扫描量热法(DSC)。进一步对制备的多孔支架进行了孔隙率测定、吸溶剂性能和不同温度下的吸水性能表征。与PLA/ HFIP支架(75%)相比,PLA/ CF支架具有更高的孔隙率(93%)和增强的吸水能力(220%)。与PLA/ HFIP和PLA/ CF支架相比,PLA/ DCM支架表现出更高的热稳定性。
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引用次数: 9
Modulation of Chondrocyte Phenotype by Bioreactor Assisted Static Compression in a 3D Polymeric Scaffold with Potential Implications to Functional Cartilage Tissue Engineering 生物反应器辅助静态压缩三维聚合物支架对软骨细胞表型的调节及其对功能性软骨组织工程的潜在影响
Pub Date : 2015-01-01 DOI: 10.4172/2157-7552.1000148
P. Nair, N. Remya
Functional cartilage tissue engineering aims at augmenting the regeneration process of chondrocyte seeded three dimensional scaffolds by application of external biomechanical stimuli. The effect of static compressive stimuli in modulating the phenotype of chondrocytes cultured in porous scaffolds using a bioreactor is being investigated in the present study. Chondrocytes were seeded in porous Poly (vinyl) alcohol-Poly capro lactone scaffold and was subjected to static unconfined compressive strain of 10% for 1h everyday for a period of 7days using a bioreactor. After culture period, chondrogenic phenotype of seeded cells was assessed by live dead assay, biochemical histological and real time PCR analysis. Bioreactor seems to be a promising tool in delivering the desired biomechanical stimuli to the cell seeded constructs. However, application of biomechanical stimuli in the form of static compression doesn’t seem beneficial as it modulates chondrogenic phenotype by reverting to a fibroblastic morphology with the secretion of collagen type 1 extra cellular matrix molecules
功能性软骨组织工程旨在利用外部生物力学刺激增强软骨细胞植入的三维支架的再生过程。静态压缩刺激对生物反应器中多孔支架中培养的软骨细胞表型的调节作用正在研究中。将软骨细胞植入多孔聚(乙烯)醇-聚卡己内酯支架中,在生物反应器中每天承受10%的静态无侧限压缩应变1h,持续7天。培养期结束后,采用活死法、生化组织学和实时PCR分析评估种子细胞的软骨表型。生物反应器似乎是一个很有前途的工具,以提供所需的生物力学刺激细胞种子结构。然而,以静态压缩的形式应用生物力学刺激似乎并不有益,因为它通过恢复成纤维细胞形态并分泌1型胶原蛋白外基质分子来调节软骨形成表型
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引用次数: 2
CD133/EpCAM Cancer Stem Cell Markers of Tumour Stage in Colorectal Cancer Cells CD133/EpCAM肿瘤干细胞标志物在结直肠癌细胞中的肿瘤分期研究
Pub Date : 2015-01-01 DOI: 10.4172/2157-7552.1000143
B. Milner, C. Penny, V. Gibbon, P. Kay, P. Ruff
In solid tumours, a discreet population of tumour associated cancer stem cells (CSCs) are proposed to drive and sustain tumour development and be responsible for tumour relapse. Colorectal cancer stem cells express cellspecific surface markers, including amongst others, CD133, EpCAM, CD44, CD166, and CD94f. In the present study, we aimed to characterisecellpopulations in the human colon adenocarcinoma cell lines, SW1116, HT29 and DLD1, expressing both CSC markers CD133 and EpCAM. These cell lines represent early, mid and late stages of colorectal tumours, respectively. Up to 107 SW1116, HT29 and DLD1 cells, co-stained with anti-CD133 and anti-EpCAM, were evaluated using flow cytometry. We report here progressive increasing proportions of cells coexpressing the CD133/EpCAM epitopes in the respective cell lines. In the SW1116 cell line, 2.42 ± 0.20 percent of cells were CD133+EpCAM+, in the HT29 cell line, 5.13 ± 0.17 percent of cells were CD133+EpCAM+, and in the DLD1 cell line, 10.30 ± 0.2 percent of cells were CD133+EpCAM+. These data suggest the frequency of CD133/ EpCAM marker expression may be associated with tumour stage and aggression.
在实体肿瘤中,肿瘤相关的肿瘤干细胞(CSCs)被认为驱动和维持肿瘤的发展,并负责肿瘤复发。结直肠癌干细胞表达细胞特异性表面标志物,包括CD133、EpCAM、CD44、CD166和CD94f等。在本研究中,我们旨在对表达CSC标记物CD133和EpCAM的人结肠癌细胞系SW1116、HT29和DLD1的细胞群进行表征。这些细胞系分别代表结直肠肿瘤的早期、中期和晚期。采用流式细胞术对107个SW1116、HT29和DLD1细胞进行抗cd133和抗epcam联合染色。我们在此报告了在各自细胞系中共表达CD133/EpCAM表位的细胞比例的逐渐增加。SW1116细胞系中CD133+EpCAM+细胞占2.42±0.20%,HT29细胞系中CD133+EpCAM+细胞占5.13±0.17%,DLD1细胞系中CD133+EpCAM+细胞占10.30±0.2%。这些数据提示CD133/ EpCAM标记物表达频率可能与肿瘤分期和侵袭性有关。
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引用次数: 5
期刊
Journal of Tissue Science and Engineering
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