Name: alginate hydrogels for three-dimensional organ culture of ovaries and oviducts
Uploaded: 2011-06-20T12:00:00
Description: Watch this Scientific Journal Video about Alginate Hydrogels for Three-Dimensional Organ Culture of Ovaries and Oviducts at JoVE.com

支持这项工作是由卵巢癌研究基金赠款LT/UIC/01.2011]，UIC的临床和转化科学，UIC的癌症中心，国家卫生部授予C06RR15482研究院的中心。 通过AAALAC认证的规定下批准的动物护理和使用委员会在UIC动物保健协议的准则和法规的规定进行了动物实验。这个项目的动物被安置在伊利诺伊州芝加哥大学的生物资源实验室（BRL）阻隔室。的BRL有一个充分的兽医人员，监控动物每天至少两次，并提供动物保健咨询。

1。卵巢解剖和组织隔离<ol><li>准备的海藻酸钠0.5％（W / V）的解决方案，准备从该协议的修改后的版本，描述以前5，使用无菌PBS和加热到37 ° C。混合倒置或摇晃，但不旋涡。绘制成1 ml注射器，海藻酸钠与25 G针，并保持在37 ° C。 </li><li> 10mm的无菌氯化钙溶液准备组织封装后交联海藻酸钠和温暖至37 ° C。 </li><li>准备1毫升培养液（αMEM+ 5个单位的青霉素和5微克/ ml链霉素），每口?...

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利用海藻酸钠水凝胶立体器官培养系统的发展代表了通用的方法，从多种生物体分析许多不同的组织类型。使用三维文化，可以扩展到组织工程和再生医学等领域，因为它提供了一个支架上的细胞能够增长11。目前，我们的实验室正处于初步研究使用人类卵巢和输卵管组织，但是，人类和非人类灵长类动物的毛囊的文化已被描述12,13。 利用海藻酸钠水凝胶的文...

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th><th>评论（可选） </th></tr><tr><td> Leibovitz的L - 15培养基</td><td> Gibco公司</td><td> 11415 </td><td></td></tr><tr><td> αMEM中等</td><td> Gibco公司</td><td> 12000-022 </td><td></td></tr><tr><td>海藻酸钠</td><td> Novamatrix </td><td></td><td></td></tr><tr><td>钠cacodylate </td><td>西格玛爱秩序</td><td> C0250 </td><td></td></tr><tr><td>聚丙烯网状纤维</td><td>麦克马斯特卡尔</td><td></td><td> 45“宽辊; 0.0041”开幕</td></tr><tr><td>脂质体2000 </td><td> Invitrogen公司</td><td> 11668 </td><td></td></tr><tr><td>青霉素/链霉素</td><td> Gibco公司</td><td> 15070-063 </td><td></td></tr><tr><td>褐藻胶裂解酶</td><td>西格玛爱秩序</td><td> A1603 </td><td></td></tr><tr><td> A1603 </td><td>西格玛爱秩序</td><td> C9697 </td><td>从梭菌histolyticum </td></tr><tr><td>牛血清白蛋白</td><td> MP生物医学</td><td> 103700 </td><td></td></tr><tr><td>胎牛血清</td><td> Gibco公司</td><td> Gibco公司</td><td></td></tr><tr><td>它的解</td><td>罗氏公司</td><td> 11074547001 </td><td>胰岛素/转/亚硒酸钠</td></tr><tr><td>细胞角蛋白8抗体</td><td>发展研究杂交瘤细胞银行</td><td> TROMA - 1 </td><td>在1:200的使用</td></tr><tr><td> DAPI </td><td>媒介实验室</td><td> H1200 </td><td></td></tr><tr><td>溴脱氧尿嘧啶核苷</td><td>西格玛</td><td> B50002 - 1G </td><td>终浓度10μM </td></tr></tbody></table>

alginate hydrogels for three-dimensional organ culture of ovaries and oviducts

卵巢癌是导致妇女因癌症死亡的第五，并已在美国的1 63％的死亡率。卵巢癌的起源的细胞类型仍然是有问题的，可能是卵巢表面上皮（OSE），或输卵管纤毛2,3远端上皮。培养原代培养的正常细胞在体外将帮助科学家模型的具体变化可能导致卵巢癌，在不同的上皮细胞，从而明确地确定起源的细胞类型。这将允许更精确的生物标志物，具有组织特异性基因的变化的动物模型，并针对本病的更好的预防策略的发展。 保持在海藻酸钠水凝胶中的正常细胞，促进细胞的体外培养中短期内其三维的背景下，允许引进的质粒DNA，RNA干扰，小分子。培养机关，从战略的削减使用手术刀，几种文化，从单一的器官可以产生，从单一的动物实验次数增加派生件。这些削减导致扩散的OSE，这是与卵巢癌的形成相关的排卵模型的各个方面。使用此方法可以培养的细胞类型，如OSE中不生长在塑料表面，并促进正常细胞过程中或在癌症形成4的最早事件进行调查。 海藻酸钠水凝胶可用于支持许多类型的组织5增长。海藻酸钠是一种线性多糖的重复单位，对β- D - mannuronic酸和α- L - guluronic酸可与钙离子交联，导致在一个温和的凝胶行动，不破坏组织6,7组成。像其他三维细胞，如基质胶文化矩阵，海藻酸钠为组织提供机械支持;但是，蛋白质不与海藻酸钠矩阵反应，并因此作为一种人工合成的细胞外基质，不会启动细胞信号5的海藻酸钠功能。海藻酸钠水凝胶漂浮在标准细胞培养液中中， 并支持在体外培养的组织生长的体系结构。 一种方法是提交到海藻酸钠水凝胶，可以在长达两个星期的文化保持的卵巢和输卵管器官件的制备，分离，嵌入。器官培养的蛋白质和RNA样品分析细胞酶的释放，也被描述。最后，主要的细胞类型增长是可能的，没有从小鼠遗传永生，并允许使用基因敲除和转基因小鼠的调查。

Watch this Scientific Journal Video about Alginate Hydrogels for Three-Dimensional Organ Culture of Ovaries and Oviducts at JoVE.com

Alginate Hydrogels for Three-Dimensional Organ Culture of Ovaries and Oviducts

正常细胞在其三维背景的文化代表了一种替代方法研究所需的细胞转化和肿瘤的早期事件。这种方法是用来种植正常的卵巢和输卵管细胞研究在卵巢癌的形成早期事件。

卵巢和输卵管的三维器官培养的海藻酸钙凝胶

Ovarian cancer is the fifth leading cause of cancer deaths in women and has a 63% mortality rate in the United States1. The cell type of origin for ...

Research

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Bioengineering

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平面和立体印刷的导电油墨

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分层海藻酸钠构造：一个异质细胞群的共培养平台

Many load bearing tissues possess structurally and functionally distinct regions, typically accompanied by different cell phenotypes with differential ...

Light-mediated Formation and Patterning of Hydrogels for Cell Culture Applications

用于细胞培养应用光介导的形成和构图的水凝胶

Click chemistries have been investigated for use in numerous biomaterials applications, including drug delivery, tissue engineering, and cell culture. In ...

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可调水凝胶由肺细胞外基质的三维细胞培养

Here we present a method for establishing multiple component cell culture hydrogels for in vitro lung cell culture. Beginning with healthy en bloc lung ...