Name: isolation of mitochondria from mouse skeletal muscle for respirometric assays
Uploaded: 2022-02-10T14:45:00
Description: Watch this Scientific Journal Video about Isolation of Mitochondria from Mouse Skeletal Muscle for Respirometric Assays at JoVE.com

我们要感谢Juan J. Tena使用均质机和CABD蛋白质组学和畜牧业设施提供技术支持。这项工作得到了西班牙教育，文化和体育部通过奖学金FPU16 / 03264到J.D.H.C，法国协会通过奖学金#22450向C.V.-G.，机构资助MDM-2016-0687（Maria de Maeztu卓越单位，CABD基因调节和形态发生系）和BFU2017-83150-P到J.J.C。Junta de Andalucía拨款P18-RT-4572，欧盟的FEDER资助计划以及西班牙科学，创新和大学部向P.N.授予RED2018-102576-T。

根据西班牙皇家法令53/2013，欧洲指令2010/63 / EU和其他相关指南，使用巴勃罗·德·奥拉维德大学伦理委员会（西班牙塞维利亚;协议24/04/2018/056和12/03/2021/033）批准的协议进行小鼠外壳和组织收集。
1. 为呼吸测定制备储备液、缓冲液和试剂
<ol><li>准备以下储备溶液，可以在指定温度下储存数月。在所有情况下使用超纯 H2O。<ol>
<li>将磷酸盐缓冲盐水（PBS）片...

<ol>
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A., Jackson, K. C., Khairallah, R. J., Ward, C. W., Spangenburg, E. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+mitochondrial+respiration+in+intact+single+muscle+fibers.">Measuring mitochondrial respiration in intact single muscle fibers.</a> American Journal of Physiology. Regulatory, Integrative and Comparative Physiology. 302 (6), 712-719 (2012).</li><li>Rosenblatt, J. D., Lunt, A. I., Parry, D. J., Partridge, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Culturing+satellite+cells+from+living+single+muscle+fiber+explants.">Culturing satellite cells from living single muscle fiber explants.</a> In Vitro Cellular &amp; Developmental Biology. 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(49), e2452 (2011).</li><li>Iuso, A., Repp, B., Biagosch, C., Terrile, C., Prokisch, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+mitochondrial+bioenergetics+in+isolated+mitochondria+from+various+mouse+tissues+using+Seahorse+XF96+analyzer.">Assessing mitochondrial bioenergetics in isolated mitochondria from various mouse tissues using Seahorse XF96 analyzer.</a> Methods in Molecular Biology. 1567, 217-230 (2017).</li><li>Divakaruni, A. S., Rogers, G. W., Murphy, A. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measuring+mitochondrial+function+in+permeabilized+cells+using+the+Seahorse+XF+analyzer+or+a+Clark-type+oxygen+electrode.">Measuring mitochondrial function in permeabilized cells using the Seahorse XF analyzer or a Clark-type oxygen electrode.</a> Current Protocols in Toxicology. 60, 1-16 (2014).</li><li>Das, K. C., Muniyappa, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age-dependent+mitochondrial+energy+dynamics+in+the+mice+heart:+role+of+superoxide+dismutase-2.">Age-dependent mitochondrial energy dynamics in the mice heart: role of superoxide dismutase-2.</a> Experimental Gerontology. 48 (9), 947-959 (2013).</li><li>Aw, W. C., Bajracharya, R., Towarnicki, S. G., Ballard, J. W. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+bioenergetic+functions+from+isolated+mitochondria+in+Drosophila+melanogaster.">Assessing bioenergetic functions from isolated mitochondria in Drosophila melanogaster.</a> Journal of Biological Methods. 3 (2), 42 (2016).</li><li>Sakamuri, S. S. V. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+respiratory+function+in+isolated+cardiac+mitochondria+using+Seahorse+XFe24+analyzer:+applications+for+aging+research.">Measurement of respiratory function in isolated cardiac mitochondria using Seahorse XFe24 analyzer: applications for aging research.</a> Gerontology. 40 (3), 347-356 (2018).</li><li>Boutagy, N. E., Pyne, E., Rogers, G. W., Ali, M., Hulver, M. W., Frisard, M. 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用于研究线粒体呼吸的所有方法都有其局限性;因此，选择最适合特定实验问题的方法至关重要。这项工作提供了一个更新和详细的方案，从小鼠骨骼肌中分离线粒体，以执行不同的呼吸测定以研究线粒体功能。事实上，使用基于微孔板的技术研究孤立线粒体中的线粒体生物能量对于研究组织特异性呼吸的可重复性，可靠性和复杂性是有价值的。此外，使用分离的线粒体可以控制底物的可用性，从...

线粒体是细胞中的主要代谢细胞器1。这些专门的膜封闭细胞器使用营养分子通过OXPHOS以三磷酸腺苷（ATP）的形式产生能量。该过程依赖于ETC2中一系列氧化还原反应中供体分子的电子转移。辅酶Q是唯一一种在所有细胞膜和循环脂蛋白中内源性产生的氧化还原活性脂质，具有抗氧化功能3。它是ETC的重要组成部分，将电子从NADH依赖性复合物I和FADH2依赖性复合物II转移到复合物III，尽管许多其他还原酶可以驱动线粒体CoQ还原为泛醇，作为多个细胞代谢途径中的强制性步骤4，5。
在整个过程中，在线粒体内膜上产生电化学质子梯度，由ATP合酶复合物V2转化为生物活性能量。因此，线粒体功能障碍导致无数的病理状况，主要影响具有高能量需求的组织 - 大脑，心脏和骨骼肌6，7。因此，开发准确分析线粒体生物能量学的方法以研究其在健康和疾病中的作用至关重要，特别是在骨骼肌等高能组织中。
克拉克型氧电极已被经典地用于线粒体呼吸的研究8。然而，该系统已逐渐被更高分辨率的技术所取代，基于微孔板的耗氧技术（如安捷伦海马 XF 分析仪）尤其受欢迎9。在骨骼肌领域，这些研究通常在培养细胞中进行，主要在C2C12永生化小鼠肌母细胞系或来自卫星细胞的原代培养物中进行10，11。然而，这些研究并不能完全概括 体内的情况，特别是在研究线粒体生物学和特定损伤，非遗传干预或遗传操作的组织水平功能时。
此外，由于其他因素，细胞中的呼吸测定更加复杂，包括ATP的线粒体外需求和测定底物或信号传导事件，这可能会误导对结果的解释。或者，也可以使用从肌肉中新鲜分离的单个或一束肌。然而，分离方法在技术上具有挑战性，仅适用于少数肌肉类型。在这种情况下，主要使用短指屈肌（FDB）和长指伸肌（EDL）10，12，13，尽管一些报告也描述了其他肌肉类型的使用14，15。
还报道了骨骼肌切片的生物能量分析16。这种方法的主要优点是可以研究完整的肌肉（作者表明，与分离的肌纤维相比，通过纤维切片不会干扰结果）。然而，线粒体对底物和测定抑制剂的访问是有限的，因此，只能测量几个参数16。最后，分离的线粒体也可以同样使用9，17，18，19。在这种情况下，线粒体会失去其胞质环境，这可能会影响其功能。相比之下，这种方法保证了对底物和抑制剂的访问，能够分析过多的样品类型，并且通常需要更少的材料。
本文描述了一种使用基于微孔板的呼吸测定对小鼠骨骼肌分离的线粒体进行生物能量分析的方法（图1）。特别是，详细列出了三种方案：偶联测定，CA，用于评估ETC和OXPHOS机器之间的偶联程度;电子流测定，EFA测量单个ETC配合物的活性;和BOX测定法测定线粒体β氧化能力。值得注意的是，与传统的呼吸法相比，只需要少量的样品。此处使用的隔离协议已从其他地方发布的方法进行了修改18。

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		<tr>
			<td>ADP</td>
			<td>Sigma</td>
			<td>A5285</td>
			<td>Stock at -20 &#176;C</td>
		</tr>
		<tr>
			<td>AKT antibody</td>
			<td>Cell Signaling Technology</td>
			<td>C67E7</td>
			<td>Rabbit (Host species)</td>
		</tr>
		<tr>
			<td>anti-Goat HRP</td>
			<td>Sigma</td>
			<td>401504</td>
			<td>Rabbit (Host species)</td>
		</tr>
		<tr>
			<td>anti-Mouse HRP</td>
			<td>Cell Signaling</td>
			<td>#7076</td>
			<td>Horse (Host species)</td>
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		<tr>
			<td>Antimycin A</td>
			<td>Sigma</td>
			<td>A8674</td>
			<td>Stock at -20 &#176;C</td>
		</tr>
		<tr>
			<td>anti-Rabbit HRP</td>
			<td>Cell Signaling</td>
			<td>#7074</td>
			<td>Goat (Host species)</td>
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		<tr>
			<td>Ascorbic acid</td>
			<td>Sigma</td>
			<td>A5960</td>
			<td>Stock at RT</td>
		</tr>
		<tr>
			<td>Bactin antibody</td>
			<td>Sigma</td>
			<td>MBS4-48085</td>
			<td>Goat (Host species)</td>
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		<tr>
			<td>Bio-Rad Protein Assay Kit II</td>
			<td>Bio-Rad</td>
			<td>5000002</td>
			<td>It includes 5x Bradford reagent and BSA of known concentration for the standard curve</td>
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		<tr>
			<td>BSA, fraction V, Fatty Acid-Free</td>
			<td>Calbiochem</td>
			<td>126575</td>
			<td>Stock at 4 &#176;C</td>
		</tr>
		<tr>
			<td>C tube</td>
			<td>Miltenyi Biotec</td>
			<td>130-093-237</td>
			<td>Purple lid</td>
		</tr>
		<tr>
			<td>Calnexin antibody</td>
			<td>ThermoFisher</td>
			<td>MA3-027</td>
			<td>Mouse (Host species)</td>
		</tr>
		<tr>
			<td>D-mannitol</td>
			<td>Sigma</td>
			<td>M4125</td>
			<td>Stock at RT</td>
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		<tr>
			<td>EDTA</td>
			<td>BDH</td>
			<td>280254D</td>
			<td>Stock at 4 &#176;C</td>
		</tr>
		<tr>
			<td>EGTA</td>
			<td>Sigma</td>
			<td>E-4378</td>
			<td>Stock at RT</td>
		</tr>
		<tr>
			<td>FCCP</td>
			<td>Sigma</td>
			<td>C2920</td>
			<td>Stock at -20 &#176;C</td>
		</tr>
		<tr>
			<td>gentleMACS Dissociator</td>
			<td>Miltenyi Biotec</td>
			<td>130-093-235</td>
			<td>Homogenizer</td>
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		<tr>
			<td>HEPES</td>
			<td>Sigma</td>
			<td>H3375</td>
			<td>Stock at RT</td>
		</tr>
		<tr>
			<td>HSP70 antibody</td>
			<td>Proteintech</td>
			<td>10995-1-AP</td>
			<td>Rabbit (Host species)</td>
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		<tr>
			<td>LDH-A antibody</td>
			<td>Santa Cruz Biotechnology</td>
			<td>SC27230</td>
			<td>Goat (Host species)</td>
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		<tr>
			<td>Magnesium chloride</td>
			<td>ChemCruz</td>
			<td>sc-255260A</td>
			<td>Stock at RT</td>
		</tr>
		<tr>
			<td>Malic acid</td>
			<td>Sigma</td>
			<td>P1645</td>
			<td>Stock at RT</td>
		</tr>
		<tr>
			<td>Microplate spectrophotometer</td>
			<td>BMG LABTECH GmbH</td>
			<td>POLARstar OMEGA S/N 415-0292</td>
			<td>Stock at RT</td>
		</tr>
		<tr>
			<td>Milli-Q water</td>
			<td>Millipore system</td>
			<td>F7HA17757A</td>
			<td>Ultrapure water</td>
		</tr>
		<tr>
			<td>mtTFA antibody</td>
			<td>Santa Cruz Biotechnology</td>
			<td>SC23588</td>
			<td>Goat (Host species)</td>
		</tr>
		<tr>
			<td>Na+/K+-ATPase &#945;1 antibody</td>
			<td>Novus Biologicals</td>
			<td>NB300-14755</td>
			<td>Mouse (Host species)</td>
		</tr>
		<tr>
			<td>Oligomycin</td>
			<td>Sigma</td>
			<td>O4876</td>
			<td>Stock at -20 &#176;C</td>
		</tr>
		<tr>
			<td>Palmitoyl-L-carnitine</td>
			<td>Sigma</td>
			<td>P1645</td>
			<td>Stock at -20 &#176;C</td>
		</tr>
		<tr>
			<td>PBS tablets</td>
			<td>Sigma</td>
			<td>P4417-100TAB</td>
			<td>1x stock at RT</td>
		</tr>
		<tr>
			<td>Potassium dihydrogen phosphate</td>
			<td>ChemCruz</td>
			<td>sc-203211</td>
			<td>Stock at RT</td>
		</tr>
		<tr>
			<td>Potassium hydroxide</td>
			<td>Sigma</td>
			<td>60377</td>
			<td>Stock at RT</td>
		</tr>
		<tr>
			<td>Pyruvic acid</td>
			<td>Sigma</td>
			<td>107360</td>
			<td>Stock at 4 &#176;C</td>
		</tr>
		<tr>
			<td>Rotenone</td>
			<td>Sigma</td>
			<td>R8875</td>
			<td>Stock at -20 &#176;C</td>
		</tr>
		<tr>
			<td>Seahorse XF24 mitochondrial flux analyzer</td>
			<td>Agilent Technologies</td>
			<td>420179</td>
			<td>XFe24 model</td>
		</tr>
		<tr>
			<td>Seahorse XFe24 FluxPak mini</td>
			<td>Agilent Technologies</td>
			<td>102342-100</td>
			<td>The kit includes cartridges, microplates, and calibrant solution</td>
		</tr>
		<tr>
			<td>Succinate</td>
			<td>Sigma</td>
			<td>S7626</td>
			<td>Stock at RT</td>
		</tr>
		<tr>
			<td>Sucrose</td>
			<td>Sigma</td>
			<td>S9378</td>
			<td>Stock at RT</td>
		</tr>
		<tr>
			<td>TIMM23 antibody</td>
			<td>Abcam</td>
			<td>ab230253</td>
			<td>Rabbit (Host species)</td>
		</tr>
		<tr>
			<td>TMPD</td>
			<td>Sigma</td>
			<td>T7394</td>
			<td>Stock at -20 &#176;C</td>
		</tr>
		<tr>
			<td>TOMM20 antibody</td>
			<td>Abcam</td>
			<td>ab56783</td>
			<td>Mouse (Host species)</td>
		</tr>
		<tr>
			<td>VDAC antibody</td>
			<td>Abcam</td>
			<td>ab15895</td>
			<td>Rabbit (Host species)</td>
		</tr>
	</tbody>
</table>

isolation of mitochondria from mouse skeletal muscle for respirometric assays

细胞的大部分能量是通过葡萄糖，脂肪酸和氨基酸的降解获得的，这些途径通过收敛于线粒体氧化磷酸化（OXPHOS）系统的不同途径获得，该系统响应细胞需求而调节。脂质分子辅酶Q（CoQ）通过恒定的氧化/还原循环将电子转移到电子传递链（ETC）中的复合物III，在此过程中是必不可少的。线粒体状态以及最终的细胞健康可以通过使用呼吸测定法测量ETC耗氧量来评估。这些研究通常在已培养数天的已建立或原代细胞系中进行。在这两种情况下，获得的呼吸参数可能偏离了任何给定器官或组织中的正常生理条件。
此外，从骨骼肌分离的培养单纤维的内在特性阻碍了这种类型的分析。本文提出了一种更新和详细的方案，用于分析从小鼠骨骼肌中新鲜分离的线粒体中的呼吸。我们还为流程任何阶段可能出现的潜在问题提供解决方案。这里介绍的方法可用于比较不同转基因小鼠模型中的耗氧率，并研究线粒体对药物治疗或其他因素（如衰老或性别）的反应。这是一种可行的方法来回答有关线粒体生物能量代谢和调节的关键问题。

这里提出的方案允许通过从小鼠骨骼肌中分离线粒体来对线粒体呼吸进行 体内 分析。该方法的概述如图 1 所示。从后肢解剖骨骼肌后（图2），在等渗条件下，通过连续离心使组织均质化和线粒体纯化。分离过程中获得的不同级分的纯度可以通过使用针对不同细胞器标记物的抗体进行蛋白质印迹分析。 图3A 显示了来自不同...

Watch this Scientific Journal Video about Isolation of Mitochondria from Mouse Skeletal Muscle for Respirometric Assays at JoVE.com

Isolation of Mitochondria from Mouse Skeletal Muscle for Respirometric Assays

在这里，我们描述了从小鼠骨骼肌中分离线粒体的详细方法，以及随后使用基于微孔板的呼吸测定通过耗氧率（OCR）分析呼吸。该管道可用于研究多种环境或遗传干预措施对线粒体代谢的影响。

从小鼠骨骼肌中分离线粒体用于呼吸测定

Most of the cell&#39;s energy is obtained through the degradation of glucose, fatty acids, and amino acids by different pathways that converge on the ...

这个词描述了处理小鼠骨骼肌线粒体中的呼吸分析。该协议大大减少了纯化时间，并允许同时处理多个样品。来自任何八种性别的United背景动物的线粒体可以使用该协议进行残缺，从而能够在各种实验条件下研究呼吸。该方法适用于与线粒体功能相关的所有研究领域，包括衰老，药物测试，癌症或发育。它甚至可以适应其他组织和生物体。在手术过程中，将所有材料都放在冰上，以确保所有材料都是冷的，以保护线粒体免受损伤。将每个样品三个50毫升烧杯放在冰上，在烧杯一中加入10毫升PBS，在烧杯二中加入10毫升10毫升10毫摩尔EDTA PBS，在烧杯三中加入四毫升IB1。用70%乙醇喷洒安乐死小鼠的右后肢以防止皮毛脱落，并将肢体贴在软木塞解剖板上，然后用胶带粘住左前肢。用无菌的一次性手术刀从膝盖到脚趾穿过皮肤做一个切口。用齿镊子在脚踝水平夹住皮肤，并在脚踝周围用细剪刀剪开。用细尖镊子一手将皮肤从下面的肌肉组织中移开，另一只手用齿镊子将其拉上来。从脚踝到膝盖解剖所有骨骼肌，并使用细镊子和剪刀去除胫骨前肌上的所有结缔组织。找到长指伸肌的四个远端肌腱，并将它们切开靠近它们在脚趾中的插入物。定位胫骨前远端肌腱，并将其切近踝关节下方的插入物。将胫骨前部和指长伸肌腱拉到踝关节上方，以释放松动的末端。抓住肌腱松弛的末端，并通过向上拉动肌肉来缓解肌肉远离骨骼中肌肉组织的其余部分。切开这些肌肉的近端肌腱，尽可能靠近膝盖骨。将鼠标倒置以继续进行腓肠肌和比目鱼肌提取。抓住肱二头肌、股骨和腓肠肌之间形成的口袋，并用细剪刀将这些肌肉分开，以观察腓肠肌近端肌腱。用细剪刀抓住并小心地切开远端跟腱。切除腓肠近端肌腱，用下面的比目鱼释放它。仔细解剖剩余的肌肉，并将鼠标重新固定在初始位置以解剖股四头肌。丢弃脂肪组织，然后在股四头肌和股骨之间插入细镊子，将肌肉与骨骼分开。抓住股四头肌远端肌腱，并尽可能靠近膝盖骨切割肌腱。将股四头肌向上拉，并在近端插入处用细剪刀将其释放。对左后肢重复相同的步骤。冲洗第一个烧杯中的所有肌肉，然后是两个，然后是烧杯三个。最后，用锋利的剪刀在冰上切碎烧杯三号中的所有肌肉。将悬浮液转移到C管上，紧紧关闭，然后将其倒置到均质机的套筒上。将匀浆分成两个2毫升预冷的微量离心管，离心10分钟。然后将上清液转移到新的预冷管中，再离心10分钟。将沉淀在500微升IB2中混合 将上清液转移到新的2毫升预冷微量离心管中，并将其标记为二号上清液或SN2。将最终的线粒体沉淀悬浮在200微升的再悬浮缓冲液中。快速留出10微升用于蛋白质定量，并立即向剩余的线粒体悬浮液中加入10微升10%FFA BSA以防止损伤。将浓缩的线粒体悬浮液以10，500倍G在4摄氏度下离心10分钟。将线粒体沉淀悬浮在100微升具有BSA的1X偶联测定培养基，具有BSA的1X电子流测定培养基或具有BSA的1Xβ氧化培养基中，具体取决于要执行的方案。使用相应的测定介质将这种浓缩的线粒体样品稀释至每微升0.2微克。将每孔50微升的悬浮液接种在冰上的预冷24孔微孔板中。在背景校正孔中，仅添加相应的测定培养基。将剩余的线粒体悬浮液储存在零下80摄氏度，以测定碎片纯度。在预冷的制备离心机中旋转微孔板20分钟。同时，将剩余的测定介质加热至37摄氏度。离心后，将微孔板放在工作台上五分钟以平衡。缓慢而小心地每孔加入450微升温热测定培养基，以避免线粒体脱落。将微孔板立即放入没有盖子的耗氧量测量仪器中，然后开始测量程序。确保第一步是孵育10分钟，以使微孔板升温。实验完成后，取出墨盒和微孔板，关闭仪器，然后开始分析。通过连续离心获得的不同组分的一般蛋白质谱在分离的组分中显示出不同的蛋白质群体。所有线粒体含量的存在都可以通过外线粒体膜和核样蛋白的标记物的强烈信号来证明。N级数代表原子核和未破裂的物质。SN1 或 SN2 级分中的大多数内质网、质膜或细胞质标志物突出了分离过程中获得的高纯度。在偶联和β氧化测定中观察到ADP添加后氧消耗量增加。羰基氰化物-对三氟甲氧基苯腙加成后，耗氧量达到最高水平。低呼吸控制比确保了分离线粒体的完整性。电子流动测定通过注射特定的底物和抑制剂单独或组合检查电子传递链复合物的活性。分离线粒体非常敏感。因此，组织缓解后的所有步骤都需要勤奋和仔细地进行，以保持其生存能力。实验测定可以通过关键线粒体分子的确定性序列分析或使用蓝色天然聚丙烯酰胺凝胶电泳研究螺旋复合物的组装来补充。

Research

JoVE Journal

Biology

Adult and Embryonic Skeletal Muscle Microexplant Culture and Isolation of Skeletal Muscle Stem Cells

成人和胚胎骨骼的肌肉Microexplant文化和骨骼肌肉干细胞的分离

Cultured embryonic and adult skeletal muscle cells have a number of different uses. The micro-dissected explants technique described in this chapter is a ...

科研

生物学

Mitochondrial Isolation from Skeletal Muscle

骨骼肌的线粒体分离

Mitochondria are organelles controlling the life and death of the cell. They participate in key metabolic reactions, synthesize most of the ATP, and ...

Purification of Progenitors from Skeletal Muscle

纯化的骨骼肌的先觉者

Skeletal muscle contains multiple progenitor populations of distinct embryonic origins and developmental potential. Myogenic progenitors, usually residing ...

Laser-inflicted Injury of Zebrafish Embryonic Skeletal Muscle

激光伤害的斑马鱼胚胎骨骼肌

Various experimental approaches have been used in mouse to induce muscle injury with the aim to study muscle regeneration, including myotoxin injections ...

Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle

单个肌纤维和成人骨骼肌卫星细胞的分离和培养

Muscle  regeneration in the adult is performed by resident stem cells called satellite  cells. Satellite cells are defined by  their position between the ...

Analysis of Skeletal Muscle Defects in Larval Zebrafish by Birefringence and Touch-evoke Escape Response Assays

Zebrafish (Danio rerio) have become a particularly effective tool for modeling human diseases affecting skeletal muscle, including muscular ...

Isolation of Blood-vessel-derived Multipotent Precursors from Human Skeletal Muscle

从人骨骼肌隔离血管源性多能前体

Since the discovery of mesenchymal stem/stromal cells (MSCs), the native identity and localization&#160;of MSCs have been obscured by their retrospective ...

Isolation and Functional Analysis of Mitochondria from Cultured Cells and Mouse Tissue

隔离和线粒体从培养细胞和小鼠组织功能分析

Comparison between two or more distinct groups, such as healthy vs. disease, is necessary to determine cellular status. Mitochondria are at the nexus of ...