A Microfluidic Chip for Efficient Circulating Tumor Cells Enrichment, Screening, and Single-Cell RNA Sequencing
A Microfluidic Chip for Efficient Circulating Tumor Cells Enrichment, Screening, and Single-Cell RNA Sequencing
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用于高效循环肿瘤细胞富集、筛选和单细胞 RNA 测序的微流控芯片
DOI：
10.1002/pmic.202000060
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发表时间：
2021-01-12
期刊：
影响因子：
3.4
通讯作者：
Hu, Zhiyuan
中科院分区：
文献类型：
--
作者：
Shi, Fanghao;Jia, Fei;Hu, Zhiyuan
循环肿瘤细胞（CTC）上的单细胞RNA测序证明可用于研究肿瘤异质性，转移和耐药性的机制。目前，CTC的单细胞RNA测序通常需要三个先决步骤：从全血中富集CTC，通过免疫染色和显微成像表征捕获的细胞，以及通过显微操作分离单细胞。然而，多个移液和转移步骤很容易导致稀有CTC的损失。为了解决这个问题，开发了一种新型的集成微流控芯片，用于在单细胞水平上连续富集、分离和表征CTC。并且，在同一芯片上实现单个CTC裂解。微流控芯片包括血凝块过滤、单细胞分离、鉴定和目标单细胞裂解物收集功能。通过将肿瘤细胞掺入全血中，验证了该微流控芯片可以有效地进行单细胞CTC RNA测序。该方法为单细胞CTC的RNA表达谱分析奠定了坚实的基础。
Single-cell RNA sequencing on circulating tumor cells (CTCs) proves useful to study mechanisms of tumor heterogeneity, metastasis, and drug resistance. Currently, single-cell RNA sequencing of CTCs usually takes three prerequisite steps: enrichment of CTCs from whole blood, characterization of captured cells by immunostaining and microscopic imaging, and single-cell isolation through micromanipulation. However, multiple pipetting and transferring steps can easily cause the loss of rare CTCs. To address this issue, a novel integrated microfluidic chip for sequential enrichment, isolation, and characterization of CTCs at single-cell level, is developed. And, single CTC lysis is achieved on the same chip. The microfluidic chip includes functions of blood clot filtration, single-cell isolation, identification, and target single-cell lysate collection. By spiking tumor cells into whole blood, it is validated that this microfluidic chip can effectively conduct single-cell CTCs RNA sequencing. The approach lays a solid foundation for the analysis of RNA expression profiling of single-cell CTCs.