Name: rotating cell culture systems for human cell culture: human trophoblast cells as a model
Uploaded: 2012-01-18T12:00:00
Description: Watch this Scientific Journal Video about Rotating Cell Culture Systems for Human Cell Culture: Human Trophoblast Cells as a Model at JoVE.com

这项工作是由美国国立卫生授予NIH / NICHD的HD051998（CAM）的机构的支持。

1。胶原珠的制备<ol><li>在此之前的3 - D细胞培养加载EVTs，需要准备的Cytodex - 3微载体珠： <ol><li>称取适量的实验所需的Cytodex - 3珠。该协议是适应农村信用社的10毫升船只，其中珠0.05克需要。对于一个50毫升农村信用社的船只，相应的规模。在一个50毫升的高压灭菌的锥形管，混合磷酸12mL贝科250毫克Cytodex - 3珠缓冲液（DPBS）。这一数额是足够5个农村信用社的船只。 </li></ol></li><li>确保有足够的量?...

<ol>
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这里介绍的文化技术提供具有高度侵袭性​​的EVT样细胞的调查。如今，它已被公认，由于抑制细胞反应，化学和分子线索的三个层面（心尖，基底和横向细胞表面 ）10,13在单层发生分化的损失。这种技术反映在子宫内指出入侵的EVT细胞的特点。由于程序模仿传统的单层组织培养的时间动力学，但提供了差异表达的细胞，比较分析可能被雇用。在实验中使用的细胞团可收获，在任?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td> Cytodex微载体珠</td><td> Sigma - Aldrich公司</td><td> C3275 </td><td></td></tr><tr><td>旋转细胞培养系统（农村信用社） </td><td> Synthecon </td><td>农村信用社- D </td><td>包括转子基地，电源供应器，4个单位一次性农村信用社</td></tr><tr><td>农村信用社一次性单位</td><td> Synthecon </td><td>联系Synthecon </td><td></td></tr><tr><td> 3毫升鲁尔锁提示注射器</td><td>屋宇署</td><td> 309585 </td><td></td></tr><tr><td> 10毫升宽提示血清移液器</td><td>屋宇署</td><td> 357504 </td><td></td></tr><tr><td>纪念阿尔法</td><td> Invitrogen公司</td><td> 12561-072 </td><td></td></tr><tr><td> Leibovitz的L - 15的介质，粉末</td><td> Invitrogen公司</td><td> 41300-039 </td><td></td></tr><tr><td> H 2 O，内毒素</td><td>费舍尔</td><td> MT - 25 - 055 - CM </td><td></td></tr><tr><td>碳酸氢钠</td><td> Sigma - Aldrich公司</td><td> S - 7795 </td><td></td></tr><tr><td>蛋白胨</td><td> Fisher Scientific则</td><td> BP1420 - 100 </td><td></td></tr><tr><td>果糖</td><td> Sigma - Aldrich公司</td><td> F3510 - 100 </td><td></td></tr><tr><td>半乳糖</td><td> Sigma - Aldrich公司</td><td> G5388 - 100 </td><td></td></tr><tr><td>血糖</td><td> Sigma - Aldrich公司</td><td> G7528 - 250 </td><td></td></tr><tr><td> HEPES </td><td> Invitrogen公司</td><td> 15630-080 </td><td></td></tr><tr><td> L -谷氨酰胺</td><td> Invitrogen公司</td><td> 25030 </td><td></td></tr><tr><td>胰岛素转亚硒酸钠（ITS）的</td><td> Sigma - Aldrich公司</td><td> I1884 </td><td></td></tr><tr><td>胎牛血清</td><td> Invitrogen公司</td><td> 10437 </td><td></td></tr><tr><td>青霉素链霉素</td><td> Invitrogen公司</td><td> 15140 </td><td></td></tr></tbody></table>

rotating cell culture systems for human cell culture: human trophoblast cells as a model

人类滋养细胞研究领域的艾滋病在认识复杂的环境中建立在胎盘。由于这些研究的性质， 人体内实验是不可能的。一个小学文化，外植体培养和滋养层细胞系的组合，支持我们的绒毛外滋养细胞（EVTs），这是怀孕的成功建立所必需的入侵的子宫壁和子宫螺旋动脉重塑3,4的理解。尽管从这种模型中收集的知识财富，它是接受，使用类似的EVT -细胞株在体外细胞培养模型显示改变细胞特性相比，其在体内的同行5,6时。培养细胞在旋转细胞培养系统（农村信用社）显示的EVT样细胞株的形态，表型和功能特性，更加紧密地模仿u的区别人Tero EVTs，调解入侵（如基质金属蛋白酶（MMPs））和滋养层细胞分化7,8,9的基因表达增加。圣乔治医院的胎盘细胞株- 4（SGHPL 4）（请盖伊惠特利博士和博士朱迪思卡特赖特捐赠）是一个EVT样细胞系，用于在农村信用社的测试。 农村信用社培养容器的设计是基于对器官和组织的原则，即在一个三维（3 - D）环境功能。由于在船只动态的文化条件，包括有关生理剪切条件，细胞生长在三个方面形成聚集基于自然细胞亲和力和分化成器官组织的集会10,11,12。维护流体轨道提供了一个低剪切，低动荡的环境相似，在体内发现的条件。培养细胞沉淀是通过调整旋转反驳加快农村信用社，以确保细胞恒定的自由落体。气体交换发生通过透水疏水膜生物反应器的背面。像他们的父母在体内的组织，能够应对在三个层面（即在其顶端，基底和横向表面），因为他们培养的多孔微载体珠表面上化学和分子梯度力场的细胞生长。像塑料防渗表面的二维膜生长时，细胞被剥夺了这一重要的沟通，在其基底表面。因此，空间限制的环境所施加深刻影响细胞如何从周围的微环境意识和解码信号， 从而暗示13的3 - D环境的重要作用。 我们已经使用了农村信用社工程师7,14,15,16各种人类上皮组织的生物学意义的3 - D模型。事实上，许多以前的报告中有DEMonstrated农村信用社培养细胞可以承担有关生理尚未10,17-21其他机型可能表型。总之，在农村信用社的文化代表了一种简单，重现性好，高通量平台，提供大量分化的细胞，适合各种实验操作。在下面的协议，作为一个例子使用EVTs，我们清楚地说明需要三个维度文化的贴壁细胞在农村信用社的步骤。

Watch this Scientific Journal Video about Rotating Cell Culture Systems for Human Cell Culture: Human Trophoblast Cells as a Model at JoVE.com

Rotating Cell Culture Systems for Human Cell Culture: Human Trophoblast Cells as a Model

传统的，二维细胞培养技术，往往导致与分化标志物，细胞因子和生长因子的改变的特点。旋转细胞培养系统的三维细胞培养（农村信用社）重新建立许多因素的表达，绒毛外滋养细胞系。

旋转的人类细胞培养的细胞培养系统：作为一种模式的滋养细胞

The field of human trophoblast research aids in  understanding the complex environment established during placentation. Due to the  nature of these ...

Research

JoVE Journal

Bioengineering

Microfabricated Platforms for Mechanically Dynamic Cell Culture

微机械动态细胞培养平台

The ability to systematically probe in vitro cellular response to combinations of mechanobiological stimuli for tissue engineering, drug discovery or ...

科研

生物工程

Shape Memory Polymers for Active Cell Culture

形状记忆聚合物主动细胞培养

Shape memory polymers (SMPs) are a class of "smart" materials that have the ability to change from a fixed, temporary shape to a pre-determined permanent ...

Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells (hESC) to Different Lineages

Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells hESC to Different Lineages

海藻酸钠微胶囊作为一个人类胚胎干细胞（胚胎干细胞）的繁殖和分化的3D平台，以不同的谱系

Human embryonic stem cells (hESC) are emerging as an attractive alternative source for cell replacement therapy since they can be expanded in culture ...

Three-dimensional Cell Culture Model for Measuring the Effects of Interstitial Fluid Flow on Tumor Cell Invasion

三维细胞培养模式的测量肿瘤细胞浸润间质流体流动的影响

The growth and progression of most solid tumors depend on the initial transformation of the cancer cells and their response to stroma-associated signaling ...

Cell Co-culture Patterning Using Aqueous Two-phase Systems

利用双水相系统的细胞共培养图案

Cell patterning  technologies that are fast, easy to use and affordable will be required for the  future development of high throughput cell assays, ...

In vitro Cell Culture Model for Toxic Inhaled Chemical Testing

In vitro Cell Culture Model for Toxic Inhaled Chemical Testing

对于中吸入有毒化学测试体外细胞培养模型

Cell cultures are indispensable to develop and study efficacy of therapeutic agents, prior to their use in animal models. We have the unique ability to ...

A Hormone-responsive 3D Culture Model of the Human Mammary Gland Epithelium

人类乳腺上皮细胞的激素反应3D文化模式

The process of mammary epithelial morphogenesis is influenced by hormones. The study of hormone action on the breast epithelium using 2D cultures is ...

Tunable Hydrogels from Pulmonary Extracellular Matrix for 3D Cell Culture

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 ...

Ultrathin Porated Elastic Hydrogels As a Biomimetic Basement Membrane for Dual Cell Culture

超薄 Porated 弹性水凝胶作为双细胞培养的仿生基底膜

The basement membrane is a critical component of cellular bilayers that can vary in stiffness, composition, architecture, and porosity. In vitro studies ...

Human Pluripotent Stem Cell Culture on Polyvinyl Alcohol-Co-Itaconic Acid Hydrogels with Varying Stiffness Under Xeno-Free Conditions

异条件下具有不同刚度的聚乙烯醇-钴-衣康酸水凝胶的人多能干细胞培养

The effect of physical cues, such as the stiffness of biomaterials on the proliferation and differentiation of stem cells, has been investigated by ...