Exosome Therapy for Stroke.
Exosome Therapy for Stroke.
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DOI：
10.1161/strokeaha.117.018292
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发表时间：
2018-05
期刊：
影响因子：
8.3
通讯作者：
Chopp M
中科院分区：
文献类型：
--
作者：
Chen J;Chopp M
2中风2018年5月尺寸排阻色谱法。他们发现，使用超离心分离，大多数分离的颗粒的直径落入外泌体的尺寸范围内;然而，当在4 ° C下应用1小时的超离心时，白蛋白也存在于制剂中。32尺寸排阻色谱分离显示出良好的再现性和快速囊泡纯化（< 10分钟）;然而，柱后外泌体浓缩步骤导致一些蛋白质损失，并且还导致低的外泌体回收率和降低的纯度（通过颗粒与蛋白质的比率评估）。29货车Deun等，33比较了4种外泌体提取方法：差速离心、OptiPrep密度梯度离心、ExoQuick沉淀和总外泌体分离沉淀。他们发现，超离心和OptiPrep密度梯度离心显示出比ExoQuick和总外泌体分离沉淀更好的通过CD 63-免疫-TEM测量的清洁外泌体制备物。OptiPrep密度梯度离心显示最纯的外泌体制备物。33然而，Taylor等人34指出，通过ExoQuick沉淀分离的循环外泌体产生的外泌体RNA和蛋白质的纯度和数量高于色谱法、超离心法和DynaBeads。因此，到目前为止，还没有用于外泌体分离的金标准方法，这使得实验室间的数据比较复杂化。21外泌体储存条件（如温度和持续时间）影响外泌体的治疗效用及其稳定性。与在-70 ℃和4 ℃下储存相比，在室温下储存10天时，外泌体的蛋白质和RNA含量降低。35外泌体在4 ℃下短期（7天内）稳定，在-80 ℃下储存至少90天。在-70 ℃以下储存是长期保存新鲜外泌体用于临床应用和基础研究的最有利条件。35
2 Stroke May 2018 size exclusion chromatography. They found that using ultracentrifugation isolation, the diameter of the majority of isolated particles fell into the size range of exosomes; however, albumin was also present in the preparations, when 1 hour of ultracentrifugation at 4 C was applied. 32 Size exclusion chromatography isolation showed good reproducibility and rapid vesicle purification (< 10 minutes); however, postcolumn exosome concentration steps resulted in some protein loss and also leads to low exosome recovery and reduced purity (assessed by the particle-to-protein ratio). 29 Van Deun et al, 33compared 4 methods of exosome extraction: differential ultracentrifugation, OptiPrep density gradient centrifugation, ExoQuick precipitation, and Total Exosome Isolation precipitation. They found that ultracentrifugation and OptiPrep density gradient centrifugation showed better clean exosome preparations measured by CD63-immuno-TEM than ExoQuick and Total Exosome Isolation precipitation. OptiPrep density gradient centrifugation showed the purest exosome preparations. 33 However, Taylor et al34 indicated that circulating exosomes isolated by ExoQuick precipitation produces exosomal RNA and protein with greater purity and quantity than chromatography, ultracentrifugation, and DynaBeads. Therefore, to date, there is no gold-standard method for exosome isolation, which complicates interlaboratory comparisons of data. 21 Exosome storage conditions (such as temperature and duration) influence the therapeutic utility of exosomes and their stability. Protein and RNA content of exosomes decrease at 10 days of room temperature storage compared with storage at− 70 and 4 C. 35 Exosomes are stable at 4 C for short term (within 7 days) and when stored at− 80 C for at least 90 days. 25 Storage at below− 70 C is the most favorable condition for long-term preservation of fresh exosomes for clinical application and basic research. 35