Name: molecular imaging to target transplanted muscle progenitor cells
Uploaded: 2013-03-27T08:54:00
Description: Watch this Scientific Journal Video about Molecular Imaging to Target Transplanted Muscle Progenitor Cells at JoVE.com

作者要感谢Sanjiv甘比尔的fluc/mrfp/sr39tk报告基因的礼物。这项工作是由加拿大干细胞网络（SCN），和杰西之旅基金会。

1。 C2C12成肌细胞的维护和传播<ol><li>板C2C12细胞在75cm 2的烧瓶中，并在高葡萄糖Dulbecco改良的Eagle的血清（HG-DMEM）中至最终浓度为10％的牛胎儿血清（FBS）补充与维持细胞。不要让细胞变得汇合在任何时间，因为这会消耗成肌人口。应该改变每一天。 注：总是温暖的介质中，以37°C的水洗澡前使用。 </li><li>当成肌细胞成为约80％汇合时，到新的烧瓶中的传代细胞。吸液培养基...

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在这项研究中，我们已经描述了一个快速和可靠的分子成像，记者基因的方法，以非侵入性的目标，成肌细胞/多用途储值卡的移植。虽然这项研究表明， 短期的本地化移植的多用途储值卡，其实很容易地应用于细胞移植的纵向评估，通过植入的细胞，细胞的方式 ，有针对性的通过bioluminescense成像（BLI），稳定表达报道基因。为此，本集团已怀有统一的报告基因的转基因小鼠品系。?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr><td> C2C12成肌细胞</td><td> ATCC </td><td></td><td></td></tr><tr><td> Dulbecco改良的Eagle培养基</td><td> Life Technologies公司</td><td> 12800-017 </td><td></td></tr><tr><td>胎牛血清</td><td> Life Technologies公司</td><td> 12483020 </td><td></td></tr><tr><td> 0.25％胰蛋白酶-EDTA </td><td> Life Technologies公司</td><td> 25200-72 </td><td></td></tr><tr><td> Hanks平衡盐溶液</td><td> Sigma Aldrich公司</td><td> H6648 </td><td></td></tr><tr><td>脂质体2000 </td><td> Life Technologies公司</td><td> 11668-019 </td><td></td></tr><tr><td>尼康的Eclipse TE2000-5 </td><td>尼康仪器公司</td><td></td><td></td></tr><tr><td> xenolight D-荧光素</td><td>珀金埃尔默</td><td> 122796 </td><td>在PBS中的40毫克/毫升</td></tr></tbody></table>

molecular imaging to target transplanted muscle progenitor cells

杜氏进行性肌营养不良症（DMD）是一种严重的遗传性神经肌肉疾病，影响1，在3500名男生，其特点是渐进性肌肉变性1，2。患者，有能力的居民肌肉卫星细胞（SC）的再生受损的肌纤维变得越来越低效4。因此，移植的肌肉祖细胞器（MPC）/成肌细胞从健康到DMD是一种很有前途的治疗方法。利用干细胞治疗的主要限制因素，然而，植入的细胞，长期监测和评价其有效性是缺乏可靠的成像技术。在这里，我们描述了一种非侵入性的，实时的方法来评价成肌细胞移植的成功。该方法利用了一个统一的融合的基因组成的报告基因（萤火虫荧光素酶[ 涨落 ]，单体红色荧光蛋白[MRFP]和SR39胸苷激酶[sr39tk基因 ]），其expre裂变可以与不同的成像模式9，10成像。多种成像模式，包括正电子发射断层扫描（PET），单光子发射计算机断层显像（SPECT），磁共振成像（MRI），光学成像，和高频率的三维超声现已有售，每一个具有独特的优势和局限性11。生物发光成像（BLI）的研究，例如，具有的优点是相对低的成本和高吞吐量。这是因为这个原因，在本研究中，我们利用鼠标后的萤火虫荧光素酶（ 涨落 ）报告基因序列内包含为短期的本地化的可行的C2C12细胞的融合基因和生物发光成像（BLI）植入到模型的DMD（肌肉萎缩症的X染色体上[MDX]鼠标）12-14。更重要的是，BLI为我们提供了一个方法来检查标记的多用途储值卡植入后的动力学，将是有益的跟踪多次在T细胞IME和下面的迁移。我们的记者基因的方法还使我们能够合并在一个单一的生物体内的多种成像模式，断层性质，精细空间分辨能力扩展到较大 ​​的动物和人类的10,11，，PET将形成未来工作的基础上，我们建议可以方便快速的翻译方法在细胞的临床前模型和临床应用开发。

当50-60％汇合，C2C12细胞瞬时转染的与上述融合报告基因构建体组成的萤火虫荧光素酶[ 涨落 ]，单体红色荧光蛋白[MRFP]和SR39胸苷激酶[sr39tk基因 ]（ 图1A）。计算转染效率，通过荧光显微镜（ 图1B中的C），在我们的记者构造使得的MRFP序列使用。不影响细胞的生存能力，通过标记与BLI基板上，D-荧光素（ 图1D）。转染后，约100,000的MRFP?...

Watch this Scientific Journal Video about Molecular Imaging to Target Transplanted Muscle Progenitor Cells at JoVE.com

Molecular Imaging to Target Transplanted Muscle Progenitor Cells

非侵入性的方法来评估成肌细胞移植的成功。该方法利用一个统一的融合的记者基因组成的基因的表达，可以被不同的成像方式成像。在这里，我们使用<em&gt;的波动</em&gt;报告基因序列到靶细胞通过生物发光成像。

目标移植的肌肉祖细胞的分子成像

Duchenne muscular dystrophy (DMD) is a severe genetic neuromuscular disorder that affects 1 in 3,500 boys, and is characterized by progressive muscle ...

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