Maximizing exosome colloidal stability following electroporation.
Maximizing exosome colloidal stability following electroporation.
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DOI：
10.1016/j.ab.2013.12.001
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发表时间：
2014-03-01
影响因子：
2.9
通讯作者：
Wickline SA
中科院分区：
文献类型：
--
作者：
Hood JL;Scott MJ;Wickline SA
用于诊断和治疗目的的基于外泌体的半合成纳米囊泡的开发需要用货物装载外泌体的新方法。电穿孔先前已用于用RNA加载外来体。然而，尚未考虑电穿孔后外泌体胶体稳定性的研究。在此，我们报告了一种独特的海藻糖脉冲培养基（TPM）的发展，最大限度地减少电穿孔后外泌体聚集。采用动态光散射（DLS）和RNA吸光度来确定与普通PBS脉冲培养基或蔗糖脉冲培养基（SPM）相比，TPM中电穿孔后外泌体聚集和电提取的程度。在电穿孔后使用TPM解聚黑素瘤外来体取决于外来体浓度和电场强度。对于同源B16黑素瘤和异质人血清来源的外来体群体，TPM使电穿孔后的外来体分散最大化。此外，TPM能够用5nm超顺磁性氧化铁纳米颗粒（SPION 5）重负载黑色素瘤外泌体，同时保持原始外泌体大小并使外泌体聚集最小化，如通过透射电子显微镜所证明的。用SPION 5装载外泌体增加了蔗糖梯度上的外泌体密度。这提供了一种简单的、无标记的方法来富集外源修饰的外泌体，并引入了MRI驱动的治疗诊断外泌体体内研究的潜力。
Development of exosome based semi-synthetic nanovesicles for diagnostic and therapeutic purposes requires novel approaches to load exosomes with cargo. Electroporation has previously been used to load exosomes with RNA. However, investigations into exosome colloidal stability following electroporation have not been considered. Herein, we report the development of a unique trehalose pulse media (TPM) that minimizes exosome aggregation following electroporation. Dynamic light scattering (DLS) and RNA absorbance were employed to determine the extent of exosome aggregation and electroextraction post electroporation in TPM compared to common PBS pulse media or sucrose pulse media (SPM). Use of TPM to disaggregate melanoma exosomes post electroporation was dependent on both exosome concentration and electric field strength. TPM maximized exosome dispersal post electroporation for both homogenous B16 melanoma and heterogeneous human serum derived populations of exosomes. Moreover, TPM enabled heavy cargo loading of melanoma exosomes with 5 nm superparamagnetic iron oxide nanoparticles (SPION5) while maintaining original exosome size and minimizing exosome aggregation as evidenced by transmission electron microscopy. Loading exosomes with SPION5 increased exosome density on sucrose gradients. This provides a simple, label free means to enrich exogenously modified exosomes and introduces the potential for MRI driven theranostic exosome investigations in vivo.
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影响因子：
46.9
作者：
Alvarez-Erviti, Lydia;Seow, Yiqi;Wood, Matthew J. A.
通讯作者：
Wood, Matthew J. A.
影响因子：
14
作者：
Safi, M.;Courtois, J.;Berret, J. -F.
通讯作者：
Berret, J. -F.
影响因子：
8.8
作者：
Taylor, DD;Gerçel-Taylor, C
通讯作者：
Gerçel-Taylor, C
影响因子：
2.7
作者：
Riteau, B;Faure, F;Rouas-Freiss, N
通讯作者：
Rouas-Freiss, N
影响因子：
4.4
作者：
Liu, CR;Yu, SH;Zhang, HG
通讯作者：
Zhang, HG