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微流控分离和血清微小囊泡的转录组分析。

Microfluidic isolation and transcriptome analysis of serum microvesicles.

机构信息

BioMEMS Resource Center, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA.

出版信息

Lab Chip. 2010 Feb 21;10(4):505-11. doi: 10.1039/b916199f. Epub 2009 Dec 8.

Abstract

Microvesicles (exosomes) shed from both normal and cancerous cells may serve as means of intercellular communication. These microvesicles carry proteins, lipids and nucleic acids derived from the host cell. Their isolation and analysis from blood samples have the potential to provide information about state and progression of malignancy and should prove of great clinical importance as biomarkers for a variety of disease states. However, current protocols for isolation of microvesicles from blood require high-speed centrifugation and filtration, which are cumbersome and time consuming. In order to take full advantage of the potential of microvesicles as biomarkers for clinical applications, faster and simpler methods of isolation will be needed. In this paper, we present an easy and rapid microfluidic immunoaffinity method to isolate microvesicles from small volumes of both serum from blood samples and conditioned medium from cells in culture. RNA of high quality can be extracted from these microvesicles providing a source of information about the genetic status of tumors to serve as biomarkers for diagnosis and prognosis of cancer.

摘要

微泡(外泌体)由正常和癌细胞脱落,可能作为细胞间通讯的一种方式。这些微泡携带源自宿主细胞的蛋白质、脂类和核酸。它们从血液样本中的分离和分析有可能提供关于恶性肿瘤状态和进展的信息,并且作为各种疾病状态的生物标志物应该具有重要的临床意义。然而,目前从血液中分离微泡的分离方案需要高速离心和过滤,这既繁琐又耗时。为了充分利用微泡作为临床应用的生物标志物的潜力,需要更快、更简单的分离方法。在本文中,我们提出了一种简单快速的微流控免疫亲和方法,用于从小体积的血清和细胞培养的条件培养基中分离微泡。可以从这些微泡中提取高质量的 RNA,为肿瘤的遗传状态提供信息,作为癌症诊断和预后的生物标志物。

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