Name: alginate microcapsule as a 3d platform for propagation and differentiation of human embryonic stem cells (hesc) to different lineages
Uploaded: 2012-03-09T12:00:00
Description: Watch this Scientific Journal Video about Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells hESC to Different Lineages at JoVE.com

这项工作是由NHMRC项目资助＃568969（PSS）和医学院，新南威尔士大学，干细胞的倡议。（KSS）支持。

下面所有的程序进行了II级生物安全柜内部使用无菌技术。在下表中列出所用的试剂和设备。 1。 1.1％的海藻酸钠的制备（W / V） <ol><li>加入纯化的海藻酸钠0.275克（高葡萄糖醛酸含量≥60％，粘度&gt; 200 MPa级小号，内毒素≤100 EU / G）在无菌的50毫升管，并添加25毫升无菌0.1％的明胶溶液，提前准备（0.5 gelatin/500毫升的Milli-Q H 2 O和由蒸压溶解）。 </li><li>涡管约30秒，?...

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	<li>Chayosumrit, M., Tuch, B., Sidhu, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alginate+microcapsule+for+propagation+and+directed+differentiation+of+hESCs+to+definitive+endoderm.">Alginate microcapsule for propagation and directed differentiation of hESCs to definitive endoderm.</a> Biomaterials. 31, 505-514 (2010).</li><li>Dean, S. K., Yulyana, Y., Williams, G., Sidhu, K. S., Tuch, B. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differentiation+of+encapsulated+embryonic+stem+cells+after+transplantation.">Differentiation of encapsulated embryonic stem cells after transplantation.</a> Transplantation. 82, 1175-1184 (2006).</li><li>Siti-Ismail, N., Bishop, A. E., Polak, J. M., Mantalaris, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+benefit+of+human+embryonic+stem+cell+encapsulation+for+prolonged+feeder-free+maintenance.">The benefit of human embryonic stem cell encapsulation for prolonged feeder-free maintenance.</a> Biomaterials. 29, 3946-3952 (2008).</li><li>Dawson, E., Mapili, G., Erickson, K., Taqvi, S., Roy, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biomaterials+for+stem+cell+differentiation.">Biomaterials for stem cell differentiation.</a> Adv. Drug Deliv. Rev. 60, 215-228 (2008).</li><li>Orive, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+encapsulation:+promise+and+progress.">Cell encapsulation: promise and progress.</a> Nat. Med. 9, 104-107 (2003).</li><li>Watanabe, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+ROCK+inhibitor+permits+survival+of+dissociated+human+embryonic+stem+cells.">A ROCK inhibitor permits survival of dissociated human embryonic stem cells.</a> Nat. Biotechnol. 25, 681-686 (2007).</li><li>Dang, S. M., Gerecht-Nir, S., Chen, J., Itskovitz-Eldor, J., Zandstra, P. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlled,+scalable+embryonic+stem+cell+differentiation+culture.">Controlled, scalable embryonic stem cell differentiation culture.</a> Stem Cells. 22, 275-282 (2004).</li><li>Drukker, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+expression+of+MHC+proteins+in+human+embryonic+stem+cells.">Characterization of the expression of MHC proteins in human embryonic stem cells.</a> Proc. Natl. Acad. Sci. U S A. 99, 9864-9869 (2002).</li><li>Calafiore, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Standard+Technical+Procedures+for+Microencapsulation+of+Human+Islets+for+Graft+into+Nonimmunosuppressed+Patients+With+Type+1+Diabetes+Mellitus.">Standard Technical Procedures for Microencapsulation of Human Islets for Graft into Nonimmunosuppressed Patients With Type 1 Diabetes Mellitus.</a> Transplantation Proceedings. 38, 1156-1157 (2006).</li><li>Cho, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Highly+efficient+and+large-scale+generation+of+functional+dopamine+neurons+from+human+embryonic+stem+cells.">Highly efficient and large-scale generation of functional dopamine neurons from human embryonic stem cells.</a> Proc. Natl. Acad. Sci. U.S.A. 105, 3392-3397 (2008).</li><li>Vazin, T., Chen, J., Lee, C. T., Amable, R., Freed, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+stromal-derived+inducing+activity+in+the+generation+of+dopaminergic+neurons+from+human+embryonic+stem+cells.">Assessment of stromal-derived inducing activity in the generation of dopaminergic neurons from human embryonic stem cells.</a> Stem Cells. 26, 1517-1525 (2008).</li></ol>

利用小鼠胚胎干细胞和胚胎干细胞的一些研究已经证明，生物材料和组织工程2,3三维培养系统的好处。我们作为一个合适的3D平台研究钡海藻酸钠相比，人类胚胎干细胞的增殖和分化，自人类胚胎干细胞表现出显着高于可行性封装比钡海藻酸钠海藻酸钙海藻酸钙微胶囊。这种文化系统还允许一个高密度细胞培养和交流的营养和氧气穿过细胞膜7。胶囊内自发分化先前已观察到的，它...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 笔记 </td></tr><tr><td>海藻酸钠（Pronova向上MVG） </td><td> NovoMatrix </td><td> 4200101 </td><td>高葡萄糖醛酸含量≥60％，粘度&gt; 200 MPa级小号，内毒素&lt;100欧/克</td></tr><tr><td>明胶</td><td> Sigma-Aldrich公司</td><td> G1890-100G </td><td></td></tr><tr><td> 0.9％的NaCl </td><td>百特医疗用品</td><td> AHF7123 </td><td></td></tr><tr><td> J1的珠发电机类型</td><td>南钢工程公司</td><td> SPA - 0447 </td><td></td></tr><tr><td>移相多注射泵</td><td>新时代热泵系统公司</td><td> NE-1000型</td><td></td></tr><tr><td> EZI流医疗流量计</td><td>吹牛系统</td><td> G0149 </td><td></td></tr><tr><td> Y型27632 </td><td>默克公司</td><td> 688000 </td><td></td></tr><tr><td>人血清白蛋白</td><td> Sigma-Aldrich公司</td><td> A4327-1G </td><td></td></tr><tr><td> accutase </td><td> Millipore公司</td><td> SCR005 </td><td></td></tr><tr><td> 14G x 2“静脉导管</td><td> Terumo公司</td><td> SR-OX1451C的</td><td></td></tr><tr><td>淘汰赛-DMEM培养液</td><td> Invitrogen公司</td><td> 10829-018 </td><td>对于SR介质（基底） </td></tr><tr><td> GlutaMAX我</td><td> Invitrogen公司</td><td> 35050-061 </td><td>对于SR培养基（2毫米） </td></tr><tr><td> Knockout血清替代品</td><td> Invitrogen公司</td><td> 10828-028 </td><td>对于SR中等（20％） </td></tr><tr><td>青霉素链霉素</td><td> Invitrogen公司</td><td> 15070063 </td><td>对于SR培养基（2.5 U / ml的） </td&gt; </tr><tr><td>胰岛素转硒区（ITS） </td><td> Invitrogen公司</td><td> 41400045 </td><td>对于SR培养基（1X） </td></tr><tr><td> β-巯基乙醇</td><td> Invitrogen公司</td><td> 21985-023 </td><td>对于SR中型（0.1毫米） </td></tr><tr><td>的MEM NEAA解决方案</td><td> Invitrogen公司</td><td> 11140-050 </td><td>对于SR中等（5毫米） </td></tr><tr><td>格拉斯哥最低基本中等</td><td> Invitrogen公司</td><td> 11710035 </td><td>对于多巴胺神经分化培养基（基底） </td></tr><tr><td> Knockout血清替代品</td><td> Invitrogen公司</td><td> 10828-028 </td><td>对于多巴胺神经分化培养基（10％） </td></tr><tr><td>丙酮酸钠</td><td> Invitrogen公司</td><td> 11360070 </td><td>对于多巴胺神经分化培养基（1毫米） </td></tr><tr><td>的MEM NEAA解决方案</td><td> Invitrogen公司</td><td> 11140-050 </td><td>对于伤残列印eural分化培养基（0.1毫米） </td></tr><tr><td> β-巯基乙醇</td><td> Invitrogen公司</td><td> 21985-023 </td><td>对于多巴胺神经分化培养基（0.1毫米） </td></tr><tr><td>刺猬（Shh） </td><td> R＆D系统</td><td> 1314-SH-025/CF </td><td> DA神经分化（100毫微克/毫升） </td></tr><tr><td>成纤维细胞生长因子8A（FGF8a） </td><td> R＆D系统</td><td> 4745-8-050 </td><td> DA神经分化（100毫微克/毫升） </td></tr></tbody></table>

alginate microcapsule as a 3d platform for propagation and differentiation of human embryonic stem cells (hesc) to different lineages

人类胚胎干细胞（胚胎干细胞）正在成为一个有吸引力的替代细胞替代治疗源，扩大在文化，因为它们可以无限期地在身体的任何类型的细胞分化。不同类型的生物材料也被用于干细胞培养提供了一个微环境，模仿的干细胞小生境1-3。后者是重要的促进提供一个三维（3D）环境中，如封装4细胞与细胞间的相互作用，细胞增殖，分化成特定的谱系，以及组织机构。 2三维文化的半渗透膜的范围内对细胞封装的原则，涉及诱捕的活细胞。这些膜允许交换横跨膜的营养物质，氧气和刺激，而从主机比胶囊孔径大的抗体和免疫细胞被排除。在这里，我们预文化派的做法和三维使用海藻酸钠微囊微环境中的人类胚胎干细胞多巴胺神经元分化。我们已经修改了培养条件，以加强封装的人类胚胎干细胞的可行性。我们先前已经表明，除了显着增强的3D平台盘绕线圈P160-Rho相关激酶（ROCK）抑制剂Y-27632和人类的胎儿成纤维细胞调节血清更换介质（HFF-CM）封装的人类胚胎干细胞的可行性在细胞中表达明确的内胚层标记基因1。我们现在已经传播到人类胚胎干细胞高效分化的多巴胺神经元用于这个3D平台。多巴胺神经细胞分化的最后阶段后，蛋白和基因表达分析表明酪氨酸羟化酶（TH）的表达增加，多巴胺神经元的标志，&gt; 2周后的100倍。我们假设，我们的3D平台，使用海藻酸钠微囊可能是有用的，以研究细胞增殖和定向分化人类胚胎干细胞，以不同的谱系。这个3D系统还允许从人类胚胎干细胞饲养层细胞的分离，并在分化过程中，也有可能在将来移植免疫隔离。

Watch this Scientific Journal Video about Alginate Microcapsule as a 3D Platform for Propagation and Differentiation of Human Embryonic Stem Cells hESC to Different Lineages at JoVE.com

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

我们已经优化了微胶囊技术作为一种有效的繁殖和胚胎干细胞分化为内胚层和多巴胺（DA）神经元的3D平台1。它还提供了一个机会，从宿​​主细胞移植过程中的免疫隔离。这个平台可以适应其他类型的细胞。

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 ...

Research

JoVE Journal

Bioengineering

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

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

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 ...

科研

生物工程

Development, Expansion, and In vivo Monitoring of Human NK Cells from Human Embryonic Stem Cells (hESCs) and Induced Pluripotent Stem Cells (iPSCs)

Development, Expansion, and In vivo Monitoring of Human NK Cells from Human Embryonic Stem Cells hESCs and Induced Pluripotent Stem Cells iPSCs

开发，扩展，<em&gt;在体内</em&gt;从人类胚胎干细胞（hESCs）监测人类NK细胞和诱导多能干细胞（iPS细胞）

We present a method for deriving natural killer (NK) cells from undifferentiated hESCs and iPSCs using a feeder-free approach. This method gives rise to ...

Ordering Single Cells and Single Embryos in 3D Confinement: A New Device for High Content Screening

订货单细胞和单胚胎的三维约束：高内涵筛选新设备

Biological cells are usually observed on flat (2D) surfaces. This condition is not physiological, and phenotypes and shapes are highly variable. Screening ...

Layered Alginate Constructs: A Platform for Co-culture of Heterogeneous Cell Populations

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 ...

Micropatterning and Assembly of 3D Microvessels

缩微和3D微血管大会

In vitro platforms to study endothelial cells and vascular biology are largely limited to 2D endothelial cell culture, flow chambers with polymer or glass ...

Single-Cell Optical Action Potential Measurement in Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes

人类诱导多能干细胞衍生心肌细胞中的单细胞光学作用潜在测量

Conventional intracellular microelectrode techniques to quantify cardiomyocyte electrophysiology are extremely complex, labor intensive, and typically ...

A Culture Method to Maintain Quiescent Human Hematopoietic Stem Cells

维持静止人类造血干细胞的文化方法

Human hematopoietic stem cells (HSCs), like other mammalian HSCs, maintain lifelong hematopoiesis in the bone marrow. HSCs remain quiescent in vivo, ...

Real-Time Intravital Multiphoton Microscopy to Visualize Focused Ultrasound and Microbubble Treatments to Increase Blood-Brain Barrier Permeability

实时活体内多光子显微镜可可视化聚焦超声和微气泡处理，以提高血脑屏障通透性

The blood-brain barrier (BBB) is a key challenge for the successful delivery of drugs to the brain. Ultrasound exposure in the presence of microbubbles ...

In vitro Induction of Human Dental Pulp Stem Cells Toward Pancreatic Lineages

In vitro Induction of Human Dental Pulp Stem Cells Toward Pancreatic Lineages

体外 人体牙浆干细胞向胰腺血统的诱导

As of 2000, the success of pancreatic islet transplantation using the Edmonton protocol to treat type I diabetes mellitus still faced some obstacles. ...

Developing 3D Organized Human Cardiac Tissue within a Microfluidic Platform

在微流体平台内开发 3D 组织人类心脏组织

The leading cause of death worldwide persists as cardiovascular disease (CVD). However, modeling the physiological and biological complexity of the heart ...