这项研究是由美国退伍军人事务部（华盛顿），补助0852048，0731201，0854430从国家科学基金会（阿灵顿，弗吉尼亚州），并从国家卫生研究院授予RO1的DK080897（马里兰州贝塞斯达的支持） 。康纳先生收到从来自比尔和梅林达·盖茨基金会（西雅图）先生爱德华·罗斯和莫妮卡莫亚博士的慷慨捐赠支持。

这里的协议描述为三个步骤，生成多层海藻酸钠微球（ 图1）。首先，海藻酸钠微球的形成（ 图2A）。此过程在下面的第1。细胞或蛋白质可以被添加到这一步的微球，以作为交付系统。下一步涉及到的微珠上permselective层的形成，并在第2条所述。最后一步涉及额外海藻酸钠层的形成，并在第3条所述。可用于外表面的珠子（ 图2B）这层形式封装，并提供治疗性分子（ 图2C）指挥系统移植后的细胞反应。 1。海藻酸钠微珠制备<ol><li>准备LVM的海藻酸钠15克溶解在1 mL的1.5％（W / V）LVM海藻酸钠溶液内层海藻酸钠溶液（25毫米HEPES缓冲液，118毫米氯化钠，氯化钾5.6毫米，2.5毫米MgCl 2的离子水，调整pH值至7.4）。上Vortexer混合海藻酸钠电源，直到完全溶解，形成一个明确的，粘性的解决方案。注：此协议描述海藻酸钠微胶囊内胰岛封装的最佳条件1的浓度和海藻酸钠微球的组成可以改变属性调整为其他应用程序（如药物释放，组织工程等）注：对于胰岛封装胰岛可以添加到海藻酸钠溶液在此之前装载在microencapsulator的一步。 </li><li>准备交联解决方案（22毫米氯化钙2 DI水）溶解CaCl 2和100毫米10毫米肝素钠DI水的缓冲和调整pH值至7.4。注：其他二价阳离子，如BR 2 +和Sr 2 +等可以用来代替的Ca 2 +，海藻酸钠凝胶d的性质取决于esired。 </li><li>增加了25号针头注射器，并调整每一侧的阀门，以确保针在空气夹克中心成立的双通道空夹克海藻酸钠microencapsulator。这一步可用于不同的表针，针对性的海藻酸钠微球的大小而定。 这一步也可以用注射器进行，应microencapsulator无法使用。加入海藻酸钠溶液的注射器，和选择的基础上，所需的微球大小的针距。 </li><li>放置一个含有10毫升的交联解决方案，直接下针的烧瓶。溶液中放置一个搅拌棒。 </li><li>以交联海藻酸钠形成微珠直接注入氯化钙溶液液滴。孵育治愈至少15分钟，不断搅拌，在交联解决方案的珠子。 </li><li>珠转移到15 mL离心管。去除残留的解决方案，执行0.2％氯化钙生理盐水2，每两分钟清洗。 </li></ol> 2。与聚-L-鸟氨酸涂层的微球<ol><li>准备3毫升生理盐水的0.1％（W / V）聚-L-鸟氨酸（巴解组织）解决方案。放置在一个旋涡的解决方案，直到完全溶解，形成了一个明确的解决方案。聚-L-赖氨酸（PLL），可以用来代替这一步，在水平相近的渗透性巴解组织。 </li><li>海藻酸钠微球转移到巴解组织解决方案。广场上的30分钟允许巴解组织足够的时间，与海藻酸钠交互，形成了阳离子，阴离子复杂的旋涡。在30分钟结束时，应该有一个明显可见的海藻酸钠微球周围的白色涂料。 </li><li>删除巴解组织解决方案，并执行0.2％的氯化钙生理盐水2，每两分钟清洗。 </li></ol> 3。创建外海藻酸钠莱呃<ol><li>准备所需的浓度，是被用来创建外层的海藻酸钠溶液。准备（1.1）以上所述的解决方案。外层的大小是由海藻酸钠的组成和使用（ 图3）浓度的影响。 </li><li>海藻酸钠微球转移到细胞的过滤。使用Kimwipe吸收多余的解决方案，以便尽可能干微珠。 </li><li>微球转移到石蜡膜表面。其他光滑的表面，如玻璃或塑料培养皿可以代替石蜡膜，如果需要的话。 </li><li>转移到海藻酸钠微球制备海藻酸钠溶液（3.1）。海藻酸钠溶液的体积应完全覆盖海藻酸钠微球。让微珠保持在45分钟的海藻酸钠溶液。 </li><li>使用吸管，去除多余的海藻酸钠溶液没有必然的微珠。 </li><li>在转移的微球到22MM 氯化钙溶液。交联海藻酸钠溶液，微珠周围形成不同外海藻酸钠层。 </li><li>执行与22毫米氯化钙生理盐水，每两分钟清洗。在显微镜下看的微球，应该允许为不同的的外海藻酸钠层看到周围形成核心微珠。 </li></ol> 4。代表结果<img src="/files/ftp_upload/3388/3388fig1.jpg" alt="图1" /> 图1为建立多层次的海藻酸钠微球的过程的示意图。从卡纳等人许可，复制。生物医学0884-2914 1.539答：十一月95（2），632-40（2010）。 <img src="/files/ftp_upload/3388/3388fig2.jpg" alt="图2" /> 图2（A）和（B）是海藻酸钠微球的相衬图像。 （a）显示了微珠后合成步骤（1.7），（二）显示了明显的步骤（3.7）完成后外海藻酸钠层。 （C）是一种荧光标记的牛血清白蛋白封装在外层蛋白FITC标记的图像。从卡纳等人许可，复制。生物医学0884-2914 1.539答：十一月95（2），632-40（2010）。 <img src="/files/ftp_upload/3388/3388fig3.jpg" alt="图3" /> 图3。外海藻酸钠层的大小可以改变的基础上使用海藻酸钠的组成和浓度。我们的研究结果表明，LVM和LVG海藻酸钠，海藻酸钠浓度的增加，最外层的大小增加，并在同等浓度比LVM的海藻酸钠，LVG海藻酸钠产生较厚的外层。从卡纳等人许可，复制。生物医学0884-2914 1.539答：十一月95（2），632-40（2010）。 <img src="/files/ftp_upload/3388/3388fig4.jpg" alt="图4"/&gt; 图4。推出FGF-1，血管生长因子蛋白变化的基础上LVM和LVG海藻酸钠使用浓度从外部海藻酸钠层。 （A）和（b）显示％释放，和（c）及（D）表示相应的质量与FGF-1的时间不同外层制剂的释放。有一阵释放展出的所有条件，在最初的5小时（A和C），低剂量的长达30天的连续释放（B和D）。从卡纳等人许可，复制。生物医学0884-2914 1.539答：月; 95（2），632-40（2010） <a href="http://www.jove.com/files/ftp_upload/3388/3388fig4large.jpg" target="_blank">点击此处查看大图</a> 。

<ol>
	<li>Darrabie, M. D., Kendall, W. F., Opara, E. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characteristics+of+Poly-L-+Ornithine-coated+alginate+microcapsules.">Characteristics of Poly-L- Ornithine-coated alginate microcapsules.</a> Biomaterials. 26, 6846-6852 (2005).</li><li>Yamagiwa, K., Kozawa, T., Ohkawa, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+alginate+composition+and+gelling+conditions+on+diffusional+and+mechanical+properties+of+calcium-alginate+gel+beads.">Effects of alginate composition and gelling conditions on diffusional and mechanical properties of calcium-alginate gel beads.</a> J. Chem. Eng. Jpn. 28, 462-467 (1995).</li><li>Amsden, B., Turner, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diffusion+characteristics+of+calcium+alginate+gels.">Diffusion characteristics of calcium alginate gels.</a> Biotechnol. Bioeng. 65, 605-610 (1999).</li><li>Uriel, S., Brey, E. M., Greisler, H. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sustained+low+levels+of+fibroblast+growth+factor-1+promote+persistent+microvascular+network+formation.">Sustained low levels of fibroblast growth factor-1 promote persistent microvascular network formation.</a> Am. J. Surg. 192, 604-609 (2006).</li><li>Moya, M. L., Lucas, S., Francis-Sedlak, M., Liu, X., Garfinkel, M. R., Huang, J. J., Cheng, M. H., Opara, E. C., Brey, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sustained+delivery+of+FGF-1+increases+vascular+density+in+comparison+to+bolus+administration.">Sustained delivery of FGF-1 increases vascular density in comparison to bolus administration.</a> Microvasc. Res. 78, 142-147 (2009).</li><li>Moya, M. L., Cheng, M. H., Huang, J. J., Francis-Sedlak, M. E., Kao, S. W., Opara, E. C., Brey, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+FGF-1+loaded+alginate+microbeads+on+neovascularization+and+adipogenesis+in+a+vascular+pedicle+model+of+adipose+tissue+engineering.">The effect of FGF-1 loaded alginate microbeads on neovascularization and adipogenesis in a vascular pedicle model of adipose tissue engineering.</a> Biomaterials. 31, 2816-2826 (2010).</li><li>Khanna, O., Moya, M. L., Greisler, H. P., Opara, E. C., Brey, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multilayered+Microcapsules+for+the+Sustained-Release+of+Angiogenic+Proteins+From+Encapsulated+Cells.">Multilayered Microcapsules for the Sustained-Release of Angiogenic Proteins From Encapsulated Cells.</a> Am. J. Surg. 200, 655-658 (2010).</li><li>Khanna, O., Moya, M. L., Opara, E. C., Brey, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+multilayered+alginate+microcapsules+for+the+sustained-release+of+fibroblast+growth+factor-1.">Synthesis of multilayered alginate microcapsules for the sustained-release of fibroblast growth factor-1.</a> J. Biomed. Mater. Res. A. 95, 632-640 (2010).</li></ol>

没有利益冲突的声明。

海藻酸钠是一种天然，从海藻中提取的酸性多糖和1,4&#39;-β-D-甘露糖醛酸（M）和α-L-古洛糖醛酸（G）的2,3单元组成。简单的凝胶发生时，二价阳离子，如Ca 2 +的Sr 2 +，或BA 2 +形成离子桥相邻海藻酸钠链的G-单体交互。海藻酸钠微球已被用来提供多种蛋白质，其中包括成纤维细胞生长因子（FGF-1），神经生长因子，白血病抑制因子，血管内皮生长因子（VEGF），包括软...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr><td> pronova超纯LVG海藻酸钠</td><td> NOVA-矩阵</td><td> 4200006 </td><td>各种海藻酸钠配方。海藻酸钠的选择影响微珠的结束，包括大小特性，力学性能，以及运输。对于一个给定的应用程序所用的组成应优化。 </td></tr><tr><td> pronova超纯LVM的海藻酸钠</td><td> NOVA-矩阵</td><td> 4200206 </td><td>各种海藻酸钠配方。海藻酸钠的选择影响微珠的结束，包括大小特性，力学性能，以及运输。对于一个给定的应用程序所用的组成应优化。 </td></tr><tr><td>聚-L-鸟氨酸盐酸盐</td><td> Sigma-Aldrich公司</td><td> P2533 </td><td></td></tr></tbody></table>

generation of alginate microspheres for biomedical applications

海藻酸钠基材料已收到相当重视医学领域的应用，由于其亲水性，生物相容性和物理结构。应用范围包括细胞封装，药物输送，干细胞培养，组织工程支架。事实上，目前正在临床试验中的小岛是封装在巴解组织涂层海藻酸钠微球作为I型糖尿病的治疗进行。然而，需要大量的胰岛由于穷人生存以下移植的疗效。局部刺​​激包裹细胞周围的微血管网络形成的能力，通过改善运输氧气，葡萄糖和其他重要的营养物质，可能会增加其可行性。成纤维细胞生长因子（FGF-1）是一种自然产生的生长因子是能刺激血管形成，改善缺血组织中的氧含量。 FGF-1的疗效SUST交付时，它是在增强ained时尚，而不是单一的大丸的管理。当地长期从胰岛封装系统的生长因子的释放，可以直接刺激血管的生长，对移植的细胞，有可能改善功能移植结果。在这篇文章中，我们概括为海藻酸钠微球在生物医学应用的准备程序。此外，我们描述的方法，我们生成多层海藻酸钠微球的发展。细胞可以被封装在海藻酸钠的内在核心，在血管生成的蛋白质外海藻酸钠层。从这个外层蛋白的释放会直接刺激当地的微血管网络的形成，对胰岛移植。

Watch this Scientific Journal Video about Generation of Alginate Microspheres for Biomedical Applications at JoVE.com

Generation of Alginate Microspheres for Biomedical Applications

在下面的章节中，我们概括为海藻酸钠微球在生物医学应用的准备程序。我们特意说明了创建多层海藻酸钠微球作为潜在的治疗1型糖尿病的细胞和蛋白质封装的双重目的的技术。

生物医学应用海藻酸钠微球的一代

Alginate-based materials have received considerable attention for biomedical applications because of their hydrophilic nature, biocompatibility, and ...

generation-of-alginate-microspheres-for-biomedical-applications

Research

JoVE Journal

Medicine

20.5K 次观看.  Illinois Institute of Technology. 在下面的章节中，我们概括为海藻酸钠微球在生物医学应用的准备程序。我们特意说明了创建多层海藻酸钠微球作为潜在的治疗1型糖尿病的细胞和蛋白质封装的双重目的的技术。

视频: Generation of Alginate Microspheres for Biomedical Applications

Generation of Comprehensive Thoracic Oncology Database - Tool for Translational Research

The Thoracic Oncology Program Database Project was created to serve as a comprehensive, verified, and accessible repository for well-annotated cancer specimens and clinical data to be available to researchers within the Thoracic Oncology Research Program. This database also captures a large volume of genomic and proteomic data obtained from various tumor tissue studies. A team of clinical and basic science researchers, a biostatistician, and a bioinformatics expert was convened to design the database. Variables of interest were clearly defined and their descriptions were written within a standard operating manual to ensure consistency of data annotation. Using a protocol for prospective tissue banking and another protocol for retrospective banking, tumor and normal tissue samples from patients consented to these protocols were collected. Clinical information such as demographics, cancer characterization, and treatment plans for these patients were abstracted and entered into an Access database. Proteomic and genomic data have been included in the database and have been linked to clinical information for patients described within the database. The data from each table were linked using the relationships function in Microsoft Access to allow the database manager to connect clinical and laboratory information during a query. The queried data can then be exported for statistical analysis and hypothesis generation.

A thoracic oncology database was developed to serve as a comprehensive repository for clinical and laboratory data for the purposes of translational research. The database will serve translational cancer researchers within the Thoracic Oncology Research Program. This database is adaptable to other cancer models, as well as other human diseases.

综合胸部肿瘤生成数据库 - 翻译研究的工具

The Thoracic Oncology Program Database Project was created to serve as a comprehensive, verified, and accessible repository for well-annotated cancer ...

科研

医学

Adaptation of Semiautomated Circulating Tumor Cell (CTC) Assays for Clinical and Preclinical Research Applications

The majority of cancer-related deaths occur subsequent to the development of metastatic disease. This highly lethal disease stage is associated with the presence of circulating tumor cells (CTCs). These rare cells have been demonstrated to be of clinical significance in metastatic breast, prostate, and colorectal cancers. The current gold standard in clinical CTC detection and enumeration is the FDA-cleared CellSearch system (CSS). This manuscript outlines the standard protocol utilized by this platform as well as two&#160;additional adapted protocols that describe the detailed process of user-defined marker optimization for protein characterization of patient CTCs and a comparable protocol for CTC capture in very low volumes of blood, using standard CSS reagents, for studying in vivo preclinical mouse models of metastasis. In addition, differences in CTC quality between healthy donor blood spiked with cells from tissue culture versus patient blood samples are highlighted. Finally, several commonly discrepant items that can lead to CTC misclassification errors are outlined. Taken together, these protocols will provide a useful resource for users of this platform interested in preclinical and clinical research pertaining to metastasis and CTCs.

Adaptation of Semiautomated Circulating Tumor Cell CTC Assays for Clinical and Preclinical Research Applications

Circulating tumor cells (CTCs) are prognostic in several metastatic cancers. This manuscript describes the gold standard CellSearch system (CSS) CTC enumeration platform and highlights common misclassification errors. In addition, two&#160;adapted protocols are described for user-defined marker characterization of CTCs and CTC enumeration in preclinical mouse models of metastasis using this technology.

半自动适应循环肿瘤细胞（CTC）分析临床和临床前研究中的应用

The majority of cancer-related deaths occur subsequent to the development of metastatic disease. This highly lethal disease stage is associated with the ...

Isolation, Cryopreservation and Culture of Human Amnion Epithelial Cells for Clinical Applications

Human amnion epithelial cells (hAECs) derived from term or pre-term amnion membranes have attracted attention from researchers and clinicians as a potential source of cells for regenerative medicine. The reason for this interest is evidence that these cells have highly multipotent differentiation ability, low immunogenicity, and anti-inflammatory functions. These properties have prompted researchers to investigate the potential of hAECs to be used to treat a variety of diseases and disorders in pre-clinical animal studies with much success.
hAECs have found widespread application for the treatment of a range of diseases and disorders. Potential clinical applications of hAECs include the treatment of stroke, multiple sclerosis, liver disease, diabetes and chronic and acute lung diseases. Progressing from pre-clinical animal studies into clinical trials requires a higher standard of quality control and safety for cell therapy products. For safety and quality control considerations, it is preferred that cell isolation protocols use animal product-free reagents. 
We have developed protocols to allow researchers to isolate, cryopreserve and culture hAECs using animal product-free reagents. The advantage of this method is that these cells can be isolated, characterized, cryopreserved and cultured without the risk of delivering potentially harmful animal pathogens to humans, while maintaining suitable cell yields, viabilities and growth potential. For researchers moving from pre-clinical animal studies to clinical trials, these methodologies will greatly accelerate regulatory approval, decrease risks and improve the quality of their therapeutic cell population.

We describe a protocol to isolate and culture human amnion epithelial cells (hAECs) using animal product-free reagents in accordance with current good manufacturing practices (cGMP) guidelines.

分离，冻存和文化的人羊膜上皮细胞的临床应用

Human amnion epithelial cells (hAECs) derived from term or pre-term amnion membranes have attracted attention from researchers and clinicians as a ...

Experimental Glaucoma Induced by Ocular Injection of Magnetic Microspheres

Progress in understanding the pathophysiology, and providing novel treatments for glaucoma is dependent on good animal models of the disease. We present here a protocol for elevating intraocular pressure (IOP) in the rat, by injecting magnetic microspheres into the anterior chamber of the eye. The use of magnetic particles allows the user to manipulate the beads into the iridocorneal angle, thus providing a very effective blockade of fluid outflow from the trabecular meshwork. This leads to long-lasting IOP rises, and eventually neuronal death in the ganglion cell layer (GCL) as well as optic nerve pathology, as seen in patients with the disease. This method is simple to perform, as it does not require machinery, specialist surgical skills, or many hours of practice to perfect. Furthermore, the pressure elevations are very robust, and reinjection of the magnetic microspheres is not usually required unlike in some other models using plastic beads. Additionally, we believe this method is suitable for adaptation for the mouse eye.

We present a method for inducing elevated intraocular pressure (IOP), by injecting magnetic microspheres into the rat eye, to model glaucoma. This leads to strong pressure rises, and extensive neuronal death. This protocol is easy to perform, does not require repeat injections, and produces stable long-lasting IOP rises.

实验诱发青光眼通过磁性微球注射眼

Progress in understanding the pathophysiology, and providing novel treatments for glaucoma is dependent on good animal models of the disease. We present ...

Three-dimensional Alginate-bead Culture of Human Pituitary Adenoma Cells

A three-dimensional culture method is described in which primary pituitary adenoma cells are grown in alginate beads. Alginate is a polymer derived from brown sea algae. Briefly, the tumor tissue is cut into small pieces and submitted to an enzymatic digestion with collagenase and trypsin. Next, a cell suspension is obtained. The tumor cell suspension is mixed with 1.2% sodium alginate and dropped into a CaCl2 solution, and the alginate/cell suspension is gelled on contact with the CaCl2 to form spherical beads. The cells embedded in the alginate beads are supplied with nutrients provided by the culture media enriched with 20% FBS. Three-dimensional culture in alginate beads maintains the viability of adenoma cells for long periods of time, up to four months. Moreover, the cells can be liberated from the alginate by washing the beads with sodium citrate and seeded on glass coverslips for further immunocytochemical analyses. The use of a cell culture model allows for the fixation and visualization of the actin cytoskeleton with minimal disorganization. In summary, alginate beads provide a reliable culture system for the maintenance of pituitary adenoma cells.

Three-dimensional culture in alginate beads, due to its simplicity and reproducibility, was chosen to maintain the pituitary adenoma cells. The procedures included an initial enzymatic digestion and mechanical dissociation of the tumor tissue, and the subsequent cell suspension was encapsulated in alginate beads.

人垂体腺瘤细胞的三维海藻酸钠珠文化

A three-dimensional culture method is described in which primary pituitary adenoma cells are grown in alginate beads. Alginate is a polymer derived from ...

Generation of Microtumors Using 3D Human Biogel Culture System and Patient-derived Glioblastoma Cells for Kinomic Profiling and Drug Response Testing

The use of patient-derived xenografts for modeling cancers has provided important insight into cancer biology and drug responsiveness. However, they are time consuming, expensive, and labor intensive. To overcome these obstacles, many research groups have turned to spheroid cultures of cancer cells. While useful, tumor spheroids or aggregates do not replicate cell-matrix interactions as found in vivo. As such, three-dimensional (3D) culture approaches utilizing an extracellular matrix scaffold provide a more realistic model system for investigation. Starting from subcutaneous or intracranial xenografts, tumor tissue is dissociated into a single cell suspension akin to cancer stem cell neurospheres. These cells are then embedded into a human-derived extracellular matrix, 3D human biogel, to generate a large number of microtumors. Interestingly, microtumors can be cultured for about a month with high viability and can be used for drug response testing using standard cytotoxicity assays such as 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and live cell imaging using Calcein-AM. Moreover, they can be analyzed via immunohistochemistry or harvested for molecular profiling, such as array-based high-throughput kinomic profiling, which is detailed here as well. 3D microtumors, thus, represent a versatile high-throughput model system that can more closely replicate in vivo tumor biology than traditional approaches.

Patient-derived xenografts of glioblastoma multiforme can be miniaturized into living microtumors using 3D human biogel culture system. This in vivo-like 3D tumor assay is suitable for drug response testing and molecular profiling, including kinomic analysis.

Microtumors使用3D人体Biogel培养体系和病人源性胶质母细胞瘤的Kinomic剖析和药物反应测试的生成

The use of patient-derived xenografts for modeling cancers has provided important insight into cancer biology and drug responsiveness. However, they are ...

Generation of Organ-conditioned Media and Applications for Studying Organ-specific Influences on Breast Cancer Metastatic Behavior

Breast cancer preferentially metastasizes to the lymph node, bone, lung, brain and liver in breast cancer patients. Previous research efforts have focused on identifying factors inherent to breast cancer cells that are responsible for this observed metastatic pattern (termed organ tropism), however much less is known about factors present within specific organs that contribute to this process. This is in part because of a lack of in vitro model systems that accurately recapitulate the organ microenvironment. To address this, an ex vivo model system has been established that allows for the study of soluble factors present within different organ microenvironments. This model consists of generating conditioned media from organs (lymph node, bone, lung, and brain) isolated from normal athymic nude mice. The model system has been validated by demonstrating that different breast cancer cell lines display cell-line specific and organ-specific malignant behavior in response to organ-conditioned media that corresponds to their in vivo metastatic potential. This model system can be used to identify and evaluate specific organ-derived soluble factors that may play a role in the metastatic behavior of breast and other types of cancer cells, including influences on growth, migration, stem-like behavior, and gene expression, as well as the identification of potential new therapeutic targets for cancer. This is the first ex vivo model system that can be used to study organ-specific metastatic behavior in detail and evaluate the role of specific organ-derived soluble factors in driving the process of cancer metastasis.

This manuscript describes an ex vivo model system comprised of organ-conditioned media derived from the lymph node, bone, lung, and brain of mice. This model system can be used to identify and study organ-derived soluble factors and their effects on the organ tropism and metastatic behavior of cancer cells.

器官空调，媒体和应用的产生对乳腺癌转移行为研究器官特异性的影响

Breast cancer preferentially metastasizes to the lymph node, bone, lung, brain and liver in breast cancer patients. Previous research efforts have focused ...

Drug Repurposing Hypothesis Generation Using the "RE:fine Drugs" System

The promise of drug repurposing is that existing drugs may be used for new disease indications in order to curb the high costs and time for approval. The goal of computational methods for drug repurposing is to enable solutions for safer, cheaper and faster drug discovery. Towards this end, we developed a novel method that integrates genetic and clinical phenotype data from large-scale GWAS and PheWAS studies with detailed drug information on the concept of transitive Drug-Gene-Disease triads. We created "RE:fine Drugs," a freely available, interactive dashboard that automates gene, disease and drug-based searches to identify drug repurposing candidates. This web-based tool supports a user-friendly interface that includes an array of advanced search and export options. Results can be prioritized in a variety of ways, including but not limited to, biomedical literature support, strength and statistical significance of GWAS and/or PheWAS associations, disease indications and molecular drug targets. Here we provide a protocol that illustrates the functionalities available in the "RE:fine Drugs" system and explores the different advanced options through a case study.

Drug Repurposing Hypothesis Generation Using the &quot;RE:fine Drugs&quot; System

Here we describe a protocol using the web-based drug repurposing hypothesis generation tool: "RE:fine Drugs." This protocol can be modified to a user's preferences at the level of the query type (gene, drug or disease) and/or the range of available advanced options.

药品再利用产生假设使用“RE：精药品”系统

The promise of drug repurposing is that existing drugs may be used for new disease indications in order to curb the high costs and time for approval. The ...

Sampling Strategies and Processing of Biobank Tissue Samples from Porcine Biomedical Models

In translational medical research, porcine models have steadily become more popular. Considering the high value of individual animals, particularly of genetically modified pig models, and the often-limited number of available animals of these models, establishment of (biobank) collections of adequately processed tissue samples suited for a broad spectrum of subsequent analyses methods, including analyses not specified at the time point of sampling, represent meaningful approaches to take full advantage of the translational value of the model. With respect to the peculiarities of porcine anatomy, comprehensive guidelines have recently been established for standardized generation of representative, high-quality samples from different porcine organs and tissues. These guidelines are essential prerequisites for the reproducibility of results and their comparability between different studies and investigators. The recording of basic data, such as organ weights and volumes, the determination of the sampling locations and of the numbers of tissue samples to be generated, as well as their orientation, size, processing and trimming directions, are relevant factors determining the generalizability and usability of the specimen for molecular, qualitative, and quantitative morphological analyses. Here, an illustrative, practical, step-by-step demonstration of the most important techniques for generation of representative, multi-purpose biobank specimen from porcine tissues is presented. The methods described here include determination of organ/tissue volumes and densities, the application of a volume-weighted systematic random sampling procedure for parenchymal organs by point-counting, determination of the extent of tissue shrinkage related to histological embedding of samples, and generation of randomly oriented samples for quantitative stereological analyses, such as isotropic uniform random (IUR) sections generated by the &#34;Orientator&#34; and &#34;Isector&#34; methods, and vertical uniform random (VUR) sections.

The practical application and performance of methods for generation of representative tissue samples of porcine animal models for a broad spectrum of downstream analyses in biobank projects are demonstrated, including volumetry, systematic random sampling, and differential processing of tissue samples for qualitative and quantitative morphologic and molecular analyses types.

猪生物医学模型生物库组织样品的取样策略与处理

In translational medical research, porcine models have steadily become more popular. Considering the high value of individual animals, particularly of ...

A Metadata Extraction Approach for Clinical Case Reports to Enable Advanced Understanding of Biomedical Concepts

Clinical case reports (CCRs) are a valuable means of sharing observations and insights in medicine. The form of these documents varies, and their content includes descriptions of numerous, novel disease presentations and treatments. Thus far, the text data within CCRs is largely unstructured, requiring significant human and computational effort to render these data useful for in-depth analysis. In this protocol, we describe methods for identifying metadata corresponding to specific biomedical concepts frequently observed within CCRs. We provide a metadata template as a guide for document annotation, recognizing that imposing structure on CCRs may be pursued by combinations of manual and automated effort. The approach presented here is appropriate for organization of concept-related text from a large literature corpus (e.g., thousands of CCRs) but may be easily adapted to facilitate more focused tasks or small sets of reports. The resulting structured text data includes sufficient semantic context to support a variety of subsequent text analysis workflows: meta-analyses to determine how to maximize CCR detail, epidemiological studies of rare diseases, and the development of models of medical language may all be made more realizable and manageable through the use of structured text data.

We present a protocol and associated metadata template for the extraction of text describing biomedical concepts in clinical case reports. The structured text values produced through this protocol can support deep analysis of thousands of clinical narratives.

一种用于临床病例报告的元数据抽取方法, 以实现对生物医学概念的深入了解

Clinical case reports (CCRs) are a valuable means of sharing observations and insights in medicine. The form of these documents varies, and their content ...