運用微流碟盤系統之密度梯度方式分離血液中轉移性癌細胞

Isolation of Metastatic Cancer Cells from Whole Blood in a Centrifugal Microfluidic Platform via Density Gradient

連偉豪(Wei-Hao Lian)

指導教授 ： 胡文聰

國立臺灣大學/工學院/應用力學研究所/碩士(2012年)

https://doi.org/10.6342/NTU.2012.10632

摘要

針對癌症病人的追蹤治療，轉移性癌細胞 (Circulating tumor cells) 是目前最具潛力的指標技術；近年來愈來愈多的研究顯示，病人血液中存在的轉移性癌細胞數量與其生存率是有相當的關係。然而在偵測技術的發展上仍非常具有挑戰性，因為其數量是極為稀少的，約莫是在數十億顆的血球細胞中尋找十顆左右的轉移性癌細胞。在過去本實驗室使用碟片系統所開發的偵測技術平台，臨床檢體的實際檢測仍無具有相當明顯的對應關係。為此，本論文的研究目的試圖將技術回歸到更直覺的方式，以腫瘤細胞本身特性而非依賴習用以免疫為基礎的方式進行分離與篩選；對於新一代的碟片平台，亦著重開發使其具有可程式自動化的流程操控。在通篇論文的闡述中，我們將有多點著墨於碟片的特性開發，包括有連續式液體注入、血球分離機制及最後蒐集液的濃縮。最後實驗的方法將呈現控制參數的最佳化，設計不同種類的癌細胞進行實驗證明此分離方式對於抗體的獨立性，並和現有常見的技術平台進行比較實驗。研究結果顯示無論種類及表現度的細胞都能至少有七成的回收率，並在細胞純度的表現上也優於其他現有的技術平台。總結而言，此研究證實了新一代的碟片具有高效率和可靠的分離機制，未來期許於病人檢體實驗上能得到不錯的臨床對應，並有機會建立起標準化的平台提供醫生作為良好的診斷工具。

關鍵字

稀少細胞 ； 轉移性癌細胞 ； 微流碟盤 ； 密度梯度 ； 細胞分離

並列摘要

Circulating tumor cell (CTC) is one of the most promising indicators for monitoring therapy in patient with metastasis cancer. A growing number of research studies are now available to shed some light on the correlation between CTCs numbers and patient’s survival. However, their cellular population is extreme rare with the peripheral blood cells that cause substantial difficulty to achieve. In our disc platform, early attempts at exploiting the detection in patient’s blood have not been very successful. Therefore, our main goal is to use a label-free density-based approach for isolation of CTCs via centrifugal separation on a disc with automated operation. This thesis focused on the features of disc, comprising of three central designs: continuous fluidic injection, separation mechanism, and concentrate enrichment. For parameters optimization, we characterized disc performance with several factors, and compared our density centrifugal disc to the conventional techniques. Result of this study showed the capture yield can achieve more than 70% in various cancer cell lines, and achieve high purity in comparison to other filtration-based approaches. To conclude, our disc platform has demonstrated high throughput and reliable microfluidics operation. This approach should be helpful to advance CTC enrichment in the fight against cancer.

並列關鍵字

Rare cell ； Circulating tumor cell ； Microfluidic disc ； Density gradient ； Cell isolation

參考文獻

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