Calcium chloride enhances the delivery of exosomes
Calcium chloride enhances the delivery of exosomes
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DOI：
10.1371/journal.pone.0220036
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发表时间：
2019-07-22
期刊：
影响因子：
3.7
通讯作者：
Joung, Boyoung
中科院分区：
文献类型：
--
作者：
Kim, Hyoeun;Kong, Ji-Young;Joung, Boyoung
外泌体作为有效的递送载体的潜力可能尚未得到改善。然而，当外泌体被开发用于治疗应用时，一种增强其递送的方法是很重要的。本研究旨在评估氯化钙（CaCl2）或其他氯化合物是否能增强外泌体对各种细胞的递送而不引起毒性。使用ExoQuick外泌体沉淀试剂盒从人血清中纯化外泌体。将分离的外泌体与浓度为100 μ M至1 mM的CaCl2混合，然后用Amicon滤池洗涤处理细胞。评估外泌体的递送效率和细胞[HEK 293（人肾细胞）和H9C2（大鼠心肌细胞）]的活力。利用PKH26标记外泌体的共聚焦显微镜观察外泌体的细胞摄取。CaCl2以剂量和治疗时间依赖的方式增加外泌体的递送。在HEK 293细胞中，CaCl2浓度为400 μ M，暴露时间为12 h，与对照相比，外泌体的递送量增加了20倍。在H9C2细胞中，CaCl2浓度为400 μ M，暴露时间为> 24 h，与对照相比，外泌体的递送量增加了> 400倍。在CaCl2浓度为1mm的情况下，两种细胞系的活力都保持不变。然而，氯化钴、氯化铜和氯化镁并没有改变两种细胞系外泌体的递送。这些结果表明，使用CaCl2处理可能是增强外泌体递送的有效方法。
Exosomes might have an unimproved potential to serve as effective delivery vehicles. However, when exosomes are developed for therapeutic applications, a method to enhance their delivery is important. This study aimed to evaluate wheather calcium chloride (CaCl2) or other chloride compounds could enhance exosome delivery to various cells without causing toxicity. Exosomes were purified from human serum by using the ExoQuick exosome precipitation kit. Isolated exosomes were mixed with CaCl2 at concentrations ranging from 100 mu M to 1 mM, and then washed using Amicon filter for treating the cells. The delivery efficiency of exosomes and the viability of the cells [HEK 293 (human kidney cells) and H9C2 (rat cardiomyocytes)] were evaluated. Cellular uptake of exosomes was observed using a confocal microscope based on PKH26 labeling of exosomes. CaCl2 increased the delivery of exosomes in a dose- and treatment time-dependent manner. In HEK 293 cells, a CaCl2 concentration of 400 mu M and exposure time of 12 h increased the delivery of exosomes by > 20 times compared with controls. In H9C2 cells, a CaCl2 concentration of 400 mu M and exposure time of > 24 h increased the delivery of exosomes by > 400 times compared with controls. The viability of both cell lines was maintained up to a CaCl2 concentration of 1 mM. However, cobalt chloride, cupric chloride, and magnesium chloride did not change the delivery of exosomes in both cell lines. These results suggest that the use of CaCl2 treatment might be a useful method for enhancing the delivery of exosomes.