基于微流控技术的循环肿瘤细胞团富集

Circulating Tumor Cell Clusters Enrichment Based on Microfluidic Technology

摘要: 近年来，癌症的死亡率逐渐升高，而肿瘤转移是造成癌症患者死亡的主要原因。极其罕见的循环肿瘤细胞团(CTC细胞团)作为高度转移性前体越来越受到重视，但它的机理目前尚不明确。因此，从癌症患者的血液中分离出可行的CTC细胞团有利于更好地研究CTC细胞团在癌症转移中的作用，并建立其作为管理疾病预后标志物的临床效用。虽然这些年来，科学家和研究员们对CTC细胞团的分离与富集有着不少的研究，但仍然有许多的局限性，比如特异性不高、纯度不高、细胞活性低等。为了解决CTC细胞团分离的一些局限性，参考了以往的文献报道，我们使用一种微流控芯片来富集捕获CTC细胞团，它依赖于CTC细胞团的不对称性，可以用于有效地将CTC细胞团与CTC细胞区分。对于在全血中的血细胞与CTC细胞团，在微流控芯片中血细胞与CTC细胞团有不同的运动轨迹。对于在缓冲液中的CTC细胞团与CTC细胞，我们做了三次平行实验，在芯片CTC细胞团回收口收集到的CTC细胞团的比例分别为71%、68%和75%，而且不影响CTC细胞团的活力，这有利于下游分子的分析。

Abstract: In recent years, the mortality rate of cancer has gradually increased, and tumor metastasis is a major cause of death in cancer patients. The extremely rare circulating tumor cell clusters (CTC clusters) are gaining more and more attention as highly metastatic precursors, but the mechanism is still unclear. Therefore, the isolation of viable CTC clusters from the blood of cancer patients is beneficial to better investigate the role of CTC clusters in cancer metastasis and to establish their clinical utility as prognostic markers for managing the disease. Although there have been many studies on CTC cluster separation and enrichment by scientists and researchers over the years, there are still many limitations, such as low specificity, low purity, and low cell activity. To address some of the limitations of CTC cluster separation, regarding previous reports in the literature, we used a microfluidic chip to enrich CTC clusters, which relies on the asymmetry of CTC clusters. It can be used to effectively distinguish between CTC clusters and CTC. For blood cells in whole blood with CTC clusters separated in the microfluidic chip, blood cells and CTC clusters have different motion trajectories. For CTC clusters and CTC in buffer, we did three parallel experiments, and the percentage of CTC clusters collected at the chip recovery port was 71%, 68% and 75%. What’s more, it did not affect the viability of CTC clusters, which facilitated the analysis of downstream molecules.