Name: mechanical stimulation of chondrocyte-agarose hydrogels
Uploaded: 2012-10-27T19:00:00
Description: Watch this Scientific Journal Video about Mechanical Stimulation of Chondrocyte-agarose Hydrogels at JoVE.com

1。主要关节软骨细胞的分离收获10-15全层动物关节的关节面的软骨片，从（ 如掌趾关节，骨骼发育成熟的奶牛从本地屠宰场获得）。 <ol><li>将软骨片在100毫米的陪替氏培养皿和孵育在20毫升0.5％的蛋白酶的Ham F-12（w / v的）为2小时，在37℃下冲洗三次火腿的F-12培养基中，并用20毫升0.15％胶原酶孵育A的Ham氏F-12培养基中过夜，在37℃下</li><li>过滤细胞悬浮液通过200目筛网...

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<html>所描述的方法应用控制的机械刺激细胞种子的琼脂糖凝胶可以对软骨细胞代谢的动态压力的影响，直接调查。护环配合使用的定制试验台结构的横向约束的样品给小费，以避免潜在的问题。尽管潜在的差异，样品高度与结构，由一组人员死加权加载压板固定使用，确保直接接触。放射性标记刺激后，测量细胞的生物合成，降低污染的可能性，允许一个单一的测试平台，以用于多个应用程序的?...

<table border="1"><tbody><tr><td> 的试剂或设备的名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr><td>联队的F-12 </td><td>赛默飞世尔科技</td><td> SH3001002 </td><td></td></tr><tr><td>胶原酶A </td><td> Sigma Aldrich公司有限公司</td><td> C0130 </td><td></td></tr><tr><td>蛋白酶</td><td> Sigma Aldrich公司有限公司</td><td> P5147 </td><td></td></tr><tr><td>胎牛血清</td><td> Sigma Aldrich公司有限公司</td><td> F1051 </td><td></td></tr><tr><td>抗坏血酸</td><td> Sigma Aldrich公司有限公司</td><td> A4034 </td><td></td></tr><tr><td>抗生素/抗真菌剂</td><td> Sigma Aldrich公司有限公司</td><td> A5955 </td><td></td></tr><tr><td> HEPES </td><td> Bioshop加拿大有限公司</td><td> HEP001 </td><td></td></tr><tr><td>台盼蓝</td><td> Sigma Aldrich公司有限公司</td><td> 93595 </td><td></td></tr><tr><td>赖克特亮线 血球</td><td>豪塞尔科学</td><td> 1490 </td><td></td></tr><tr><td>定量，它PicoGreen </td><td> Invitrogen公司</td><td> P7589 </td><td></td></tr><tr><td>木瓜木瓜乳胶</td><td> Sigma Aldrich公司有限公司</td><td> P3125 </td><td></td></tr><tr><td>醋酸铵</td><td> Sigma Aldrich公司有限公司</td><td> A1542 </td><td></td></tr><tr><td> Ethyldiaminetetraacetic酸</td><td> Sigma Aldrich公司有限公司</td><td> E9884 </td><td></td></tr><tr><td> DL-二硫苏糖醇</td><td> Sigma Aldrich公司有限公司</td><td> 43819 </td><td></td></tr><tr><td>低熔点琼脂糖，第七类</td><td> Sigma Aldrich公司有限公司</td><td> A9045 </td><td></td></tr> &lt;TR&gt; <td>筛网过滤器（200） </td><td> Sigma Aldrich公司有限公司</td><td> S4145 </td><td></td></tr><tr><td> MACH-1微机械测试仪</td><td> Biomomentum公司</td><td> V500cs </td><td></td></tr><tr><td>压缩荷载跳汰机</td><td>定制</td><td></td><td>同类产品可以提供的Biomomentum公司</td></tr><tr><td>猎鹰24孔培养板</td><td>赛默飞世尔科技</td><td> B353047 </td><td></td></tr><tr><td> β-液体闪烁计数器</td><td> Beckman Coulter公司</td><td> LS6500 </td><td></td></tr><tr><td> [3H]脯氨酸</td><td> Perkin-Elmer公司</td><td> NET323005MC </td><td></td></tr><tr><td> [35]硫</td><td> Perkin-Elmer公司</td><td> NEX041005MC </td><td></td></tr></tbody></table>

mechanical stimulation of chondrocyte-agarose hydrogels

机械侮辱或退化性疾病，如骨关节炎关节软骨受到损坏时，从有限的修复能力。为了弥补这方面的不足，一些医疗干预措施已经制定出来。一种方法是复出与组织工程化软骨的受损区域，然而，组织工程通常缺乏本地软骨的生化特性和耐用性，质疑它的长期生存能力。这限制了小灶缺陷修复软骨组织工程的应用，依靠对周围组织的保护植入材料。为了改善性能的发达的组织，机械性刺激是一种流行的方法，利用增强软骨细胞外基质的合成，以及所得的机械性能的组织工程。机械性刺激，适用于力量的组织构造类似于那些有经验的体内 。这是基于的前提是机械的环境中，在某种程度上，调节的开发和维护的固有组织1,2。在软骨组织工程的机械性刺激中最常采用的形式是在生理菌株在1,3 1赫兹的频率的约5-20％的动态压缩。一些研究调查的影响，动态压缩，并已显示出它有一个积极的影响软骨细胞的代谢和生物合成，最终影响功能特性的开发组织4-8。在本文中，我们说明了该方法机械地刺激软骨细胞琼脂糖凝胶结构下的动态压缩和生物合成的变化进行分析，通过生化和放射性同位素检测。此方法也可以很容易地修改，作为一个结果，机械刺激，以评估任何可能引起的变化在细胞反应。

Watch this Scientific Journal Video about Mechanical Stimulation of Chondrocyte-agarose Hydrogels at JoVE.com

Mechanical Stimulation of Chondrocyte-agarose Hydrogels

软骨细胞的软骨细胞外基质的生物合成的，可以通过施加机械刺激的影响。该方法描述了技术应用动态压缩应变包埋软骨细胞的三维结构和诱导软骨细胞代谢的变化的评价。

机械性刺激软骨细胞琼脂糖凝胶

Articular cartilage suffers from a limited repair capacity when damaged by mechanical insult or degraded by disease, such as osteoarthritis. To remedy ...

Research

JoVE Journal

Biology

Ex vivo Mechanical Loading of Tendon

体外机械装载肌腱

Injuries to the tendon (e.g., wrist tendonitis, epicondyltis) due to overuse are common in sports activities and the workplace.  Most are associated with ...

科研

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Mechanical Stimulation of Stem Cells Using Cyclic Uniaxial Strain

采用循环单轴应变的干细胞的机械性刺激

The role of mechanical forces in the development and maintenance of biological tissues is well documented, including several mechanically regulated ...

Cellular Encapsulation in 3D Hydrogels for Tissue Engineering 

组织工程细胞在三维凝胶封装

The 3D encapsulation of cells within hydrogels represents an increasingly important and popular technique for culturing cells and towards the development ...

Live Cell Response to Mechanical Stimulation Studied by Integrated Optical and Atomic Force Microscopy

机械性刺激，集成光学和原子力显微镜研究活细胞的反应

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Mechanical Stimulation-induced Calcium Wave Propagation in Cell Monolayers: The Example of Bovine Corneal Endothelial Cells

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Preparation of DNA-crosslinked Polyacrylamide Hydrogels

DNA交联聚丙烯酰胺水凝胶的制备

Mechanobiology is an emerging scientific area that addresses the critical role of physical cues in directing cell morphology and function. For example, ...

Multifunctional, Micropipette-based Method for Incorporation And Stimulation of Bacterial Mechanosensitive Ion Channels in Droplet Interface Bilayers

多功能，微管为基础的法团细菌机械敏感离子通道与刺激的液滴界面双层膜

MscL, a large conductance mechanosensitive channel (MSC), is a ubiquitous osmolyte release valve that helps bacteria survive abrupt hypo-osmotic shocks. ...

FRET Imaging in Three-dimensional Hydrogels

FRET成像的三维水凝胶

Imaging of F&#246;rster resonance energy transfer (FRET)&#160;is a powerful tool for examining cell biology in real-time. Studies utilizing FRET commonly ...

Assessment of Dictyostelium discoideum Response to Acute Mechanical Stimulation

Assessment of Dictyostelium discoideum Response to Acute Mechanical Stimulation

对急性机械刺激的网柄反应的评估

Chemotaxis, or migration up a gradient of a chemoattractant, is the best understood mode of directed migration. Studies using social amoeba Dictyostelium ...

Proteolytically Degraded Alginate Hydrogels and Hydrophobic Microbioreactors for Porcine Oocyte Encapsulation

蛋白酶降解藻酸盐水凝胶和疏水性微生物反应器，用于猪卵母细胞封装

In reproductive biology, the biotechnology revolution that began with artificial insemination and embryo transfer technology led to the development of ...