Name: engineering skeletal muscle tissues from murine myoblast progenitor cells and application of electrical stimulation
Uploaded: 2013-03-19T12:00:00
Description: Watch this Scientific Journal Video about Engineering Skeletal Muscle Tissues from Murine Myoblast Progenitor Cells and Application of Electrical Stimulation at JoVE.com

要感谢亚斌吴培养的组织图2给出了，照片是由巴特面包车Overbeeke。 SenterNovem，授予ISO 42022的工作的财政支持。

1。小鼠成肌细胞的祖细胞或C2C12细胞培养<ol><li>隔离细胞根据最初由谢费尔和他的同事10出版和购买适于由Collins 等人 11和博南等12的协议，并在液氮中存储这些。这需要小鼠， 如之C57B1 / 6。在其他实验室所使用的替代方法， 例如李瑛等人发表在杂志的可视化实验的方法13。对于试剂和设备，被称为第7和第8页上的表。从一...

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肌肉组织工程具有巨大的潜力作为一种疾病​​模型，药物筛选，在再生医学中的使用和肉类生产。然而，这些应用程序的要求有所不同。我们选择与胶原蛋白和基质胶的组合，因为胶原蛋白可以用于细胞取向和成肌细胞前体细胞，因为需要基底膜衍生的蛋白质的存在下，如在之前的2D研究12确定。此外，纤维蛋白凝胶已在我们的实验室测试，似乎不支持C2C12肌母细胞的祖细胞，胶原蛋白?...

<table border="1"><tbody><tr><td> 的试剂的名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr><td> Matrigel的生长因子减少</td><td> Beckton和Dickinson </td><td></td><td></td></tr><tr><td> DMEM（高糖）* </td><td> Gibco公司</td><td> 42430 </td><td></td></tr><tr><td>先进的DMEM </td><td> Gibco公司</td><td> 12491 </td><td></td></tr><tr><td>马血清</td><td> Gibco公司</td><td> 65050-122 </td><td></td></tr><tr><td>胎牛血清</td><td>格雷纳</td><td> 758075 </td><td></td></tr><tr><td> 0.45微米和0.22微米针头式过滤器</td><td> Whatmann（Schleicher和Scheull） </td><td> 10462100 </td><td></td></tr><tr><td> L-谷氨酰胺</td><td> Gibco公司</td><td> 25030024 </td><td></td></tr><tR&gt; <td>青霉素/链霉素</td><td> Gibco公司</td><td> 10378016 </td><td></td></tr><tr><td>两性霉素</td><td> Gibco公司</td><td> 15290-018 </td><td></td></tr><tr><td>文化塑胶</td><td>格雷纳</td><td></td><td>包括培养瓶和吸液管</td></tr><tr><td>鸡胚提取物</td><td>美国生物</td><td> C3999 </td><td></td></tr><tr><td>巴斯德吸管* </td><td> Hilgenberg </td><td></td><td>巴氏吸管，收缩，与棉，顶端开口L：230毫米，尖端直径0.9 - 1.1毫米</td></tr><tr><td>巴斯德吸管* </td><td> Hilgenberg </td><td></td><td>巴氏吸管，收缩，与棉，顶端开口L：尖端直径230毫米，1,4 - 1,6毫米</td></tr><tr><td>巴斯德吸管</td><td> VWR </td><td> 612-1702 </td><td></td></tr><tr><td> I型胶原酶* </td><td>西格玛</td><td> C0130-16 </td><td></td></tr><tr><td> 40微米的细胞过滤* </td><td> BD猎鹰</td><td> 352340 </td><td></td></tr><tr><td> 19G针</td><td></td><td></td><td></td></tr><tr><td>弹性体</td><td>道康宁公司</td><td> 3097358-1004 </td><td>硅橡胶MDX 4-4210＃ </td></tr><tr><td>固化剂</td><td>道康宁公司</td><td></td><td>硅橡胶MDX 4-4210＃ </td></tr><tr><td>尼龙搭扣</td><td>正规商店</td><td></td><td>您可以在正规的商店购买，只能使用柔软的一面</td></tr><tr><td> Ⅰ型胶原，鼠尾</td><td> BD Biosciences公司</td><td> 3544236 </td><td></td></tr><tr><td> C-吴佩慈EP文化步行者</td><td> Ionoptix </td><td></td><td></td></tr><tr><td> 6孔培养板，电刺激</td><td>卡特莱特kton迪金森猎鹰</td><td> BD猎鹰＃353846 </td><td></td></tr><tr><td> C-DISH培养皿电极</td><td> Ionoptix </td><td></td><td></td></tr><tr><td></td><td></td><td></td><td> *所需要的隔离的细胞（1.1） ＃在同一个工具包</td></tr></tbody></table>

engineering skeletal muscle tissues from murine myoblast progenitor cells and application of electrical stimulation

工程的肌肉组织，可用于几个不同的用途，包括用于生产的组织在体外作为一种疾病模型， 如学习压力性溃疡，再生医学和肉类替代1。首次报道的3D肌肉结构已作了很多年前在该领域的先驱，是Vandenburgh和他的同事2,3。在肌肉组织工程取得的进展不仅是广大的生化因子，干细胞和祖细胞知识增益的结果，但研究人员所获得的洞察力，特别是基于物理因素发挥重要作用，在控制细胞的行为，组织的发展。国家的最先进的工程肌肉构造目前包括人口细胞的水凝胶结构。在我们的实验室中，这些通常由鼠成肌细胞的祖细胞，分离出小鼠后肢肌肉或小鼠成肌细胞系C2C12，MIXED与胶原/基底膜的混合物，并镀两个锚固点之间，模仿肌肉韧带。其他细胞可能被视为良好， 例如 ，替代的细胞系，例如L6大鼠成肌细胞4，新生儿肌源性祖细胞5，来自成年肌肉组织从其他物种如人类6甚至诱导式多能性干细胞（iPS细胞）7的细胞。细胞收缩引起的细胞的取向沿长轴的构建体8,9和文化约一个星期后的肌肉祖细胞的分化。此外，应用程序可以增强电刺激分化的过程中，在一定程度上8。由于其有限的尺寸（8×2×0.5 mm）的完整的组织可以使用共焦显微镜分析，监视例如生存能力，分化和细胞对准。根据特定的应用要求的enginee红色的肌肉组织会有所不同， 例如用于再生医学需要向上缩放的组织大小和血管，而作为一个肉替代的词条到其他物种是必要的。

最终产品将是肌肉如在图3中所示的构造。组织的大小将是约8毫米长，2毫米宽，0.5毫米厚。在分化过程中的电刺激将改变的表达，肌球蛋白重链异构体，但不大大提高的分化的方法，诱导分化培养基8，但也可以应用于电刺激可以肌肉的功能的过程结束时，判断是否，因为将能够充分​​发展的肌节的肌肉收缩时的电脉冲。 分化和成熟的也可以使用定量PCR来分析?...

Watch this Scientific Journal Video about Engineering Skeletal Muscle Tissues from Murine Myoblast Progenitor Cells and Application of Electrical Stimulation at JoVE.com

Engineering Skeletal Muscle Tissues from Murine Myoblast Progenitor Cells and Application of Electrical Stimulation

工程肌肉组织再生医学具有巨大的潜力，作为疾病模型，也可作为肉类的替代来源。在这里，我们描述的工程的肌肉构造，在这种情况下，从小鼠成肌细胞的祖细胞，通过电脉冲刺激。

工程骨骼肌组织中的小鼠成肌细胞的内皮祖细胞与应用电刺激

Engineered muscle tissues can be used for several different purposes, which include the production of tissues for use as a disease model in vitro, e.g. to ...

Research

JoVE Journal

Bioengineering

Fabrication of Micro-tissues using Modules of Collagen Gel Containing Cells

微组织使用胶原蛋白的模块的制备凝胶含有细胞

This protocol describes the fabrication of a type of micro-tissues called modules.  The module approach generates uniform, scalable and vascularized ...

科研

生物工程

Engineering Fibrin-based Tissue Constructs from Myofibroblasts and Application of Constraints and Strain to Induce Cell and Collagen Reorganization

肌成纤维细胞和约束的应用工程纤维蛋白的​​组织结构和应变诱导细胞和胶原蛋白（RE）组织

Collagen content and organization in developing collagenous tissues can be influenced by local tissue strains and tissue constraint. Tissue engineers aim ...

Electrospun Fibrous Scaffolds of Poly(glycerol-dodecanedioate) for Engineering Neural Tissues From Mouse Embryonic Stem Cells

Electrospun Fibrous Scaffolds of Polyglycerol-dodecanedioate for Engineering Neural Tissues From Mouse Embryonic Stem Cells

保利的静电纤维支架（甘油 - 十二双）工程神经组织从小鼠胚胎干细胞

For tissue engineering applications, the preparation of biodegradable and biocompatible scaffolds is the most desirable but challenging task. &#160;Among ...

Measurement of Maximum Isometric Force Generated by Permeabilized Skeletal Muscle Fibers

最大等长收缩力的测量通过透骨骼肌纤维生成

Analysis of the contractile properties of chemically skinned, or permeabilized, skeletal muscle fibers offers a powerful means by which to assess muscle ...

Cardiac Muscle-cell Based Actuator and Self-stabilizing Biorobot - PART 1

心肌肌细胞基致动器和自稳定Biorobot  - 第1部分

Biological machines often referred to as biorobots, are living cell- or tissue-based devices that are powered solely by the contractile activity of living ...

Cardiac Muscle Cell-based Actuator and Self-stabilizing Biorobot - Part 2

心肌肌细胞基致动器和自稳定Biorobot  - 第2部分

In recent years, hybrid devices that consist of a living cell or tissue component integrated with a synthetic mechanical backbone have been developed. ...

Microfluidic Bioprinting for Engineering Vascularized Tissues and Organoids

微流控生物工程血管化组织和细胞器解体

Engineering vascularized tissue constructs and organoids has been historically challenging. Here we describe a novel method based on microfluidic ...

Two Methods for Decellularization of Plant Tissues for Tissue Engineering Applications

组织工程应用中植物组织去细胞的两种方法

The autologous, synthetic, and animal-derived grafts currently used as scaffolds for tissue replacement have limitations due to low availability, poor ...

Simultaneous Electrical and Mechanical Stimulation to Enhance Cells' Cardiomyogenic Potential

同时进行机电刺激增强细胞心肌成功能的潜能

Cardiovascular diseases are the leading cause of death in developed countries. Consequently, the demand for effective cardiac cell therapies has motivated ...

Construction and Use of an Electrical Stimulation Chamber for Enhancing Osteogenic Differentiation in Mesenchymal Stem/Stromal Cells In Vitro

电刺激室的构建与应用, 促进间充质茎间质细胞体外成骨分化

Mesenchymal stem/stromal cells (MSCs) have been used extensively to promote bone healing in tissue engineering approaches. Electrical stimulation (EStim) ...