全血中單細胞回收之自動化微流系統開發

An automated, seamless microfluidic system for single cell isolation and retrieval from whole blood

李孟澤(Meng-Ze Li)

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(2014年)

https://doi.org/10.6342/NTU.2014.10290

摘要

循環腫瘤細胞於現階段被認為是一種具有檢測與診斷癌症潛力的指標。許多研究已驗證了循環腫瘤細胞的數量與患者的存活率有明顯的關係，但這種細胞本身極度稀少且異質性高，而現有大部份針對其純化之技術的靈敏度還需要被佐證的狀況下，這些循環腫瘤細胞計數的資訊目前於臨床上並不具有實質的效用。為了更加了解循環腫瘤細胞於癌症轉移的過程中所具有的臨床價值，對這些細胞進行定性分類、單細胞分析與分子分析的研究開始受到關注。本論文展示一套搭配特殊設計平台的細胞抓取系統，以幫助臨床學家與腫瘤學者更容易取得高度純化的細胞來針對循環腫瘤細胞進行更深入的研究。
我們設計並建構了一套細胞抓取系統用於取出循環腫瘤細胞。為了提高其效率，這套系統中具有特殊的設計以減少在抓取過程中對周遭非目標細胞的擾動。實驗結果顯示目標細胞的成功抓取率幾近100%且其抓取純率超越90%，遠高於原始設計。此外我們利用健康人全血混入Colo 205的實驗展示由碟盤系統初步分選至自動化細胞抓取的整合流程，並取得過半的目標細胞與幾近100%的抓取純率。總的來說，此細胞抓取系統藉著特殊設計平台的幫助已可實現初步的自動化，同時兼具高抓取回收率與抓取純率，可望能在未來成為一套有力的系統來協助循環腫瘤細胞之後端研究。

關鍵字

循環腫瘤細胞 ； 細胞抓取 ； 抓取純率 ； 自動化

並列摘要

Circulating tumor cell (CTC) now is thought to be a possible surrogate for cancer diagnosis and prognosis. Many studies have reported worse survival of patients in different cancer types was related to the higher number of detected CTCs. However, CTC count information still do not possess clinical utility to date since CTC presents high rarity and heterogeneity, yet the sensitivity of most technologies for CTC enrichment remain to be validated. To advance the understanding and clinical value of CTC in metastasis process, there has been great interest in subtyping, single cell profiling and molecular analysis of CTC. This thesis presents a cell retrieval system with a modified platform as a beneficial tool helping clinicians and oncologists to obtain highly purified cells of interest which allows them to explore deeper potential of CTC.
We designed and constructed a cell retrieval system to remove CTC, which would be extremely difficult without the special design due to flow interaction between neighboring cells during the retrieval process. Results show cell retrieval rate reached nearly 100% of target cells and the average retrieval purity was over 90%, which is much higher than the original design. Moreover, we used Colo 205 spiked into blood sample to demonstrate the seamless process from primary isolation of disk system to automatic cell retrieval which has been successfully performed with over half target cells were retrieved of a nearly 100% retrieval purity. In conclusion, the cell retrieval system with the modified platform has realized the primary automation of cell retrieving technique with high retrieval rate and high retrieval purity, which should become a powerful tool to assist CTC characterization and molecular analysis.

並列關鍵字

CTC ； cell retrieval ； retrieval purity ； automation

參考文獻

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