攜帶螢光及磁性奈米鑽石細胞的分離、鑑定與應用

Abstract

奈米鑽石是一種含碳的奈米材料，近年被重視開發於生物醫學的應用。在本篇論文中我們使用螢光磁性奈米鑽石，利用其帶有螢光與磁性，開發出一種攜帶奈米鑽石的新穎細胞株。螢光磁性奈米鑽石不會對正常HFL-1肺纖維母細胞及A549肺上皮細胞產生明顯的細胞毒性。A549細胞處理螢光磁性奈米鑽石後，隨著濃度及時間的增加，細胞內螢光強度與顆粒複雜度亦隨之升高，然而細胞大小的分布並不受螢光磁性奈米鑽石的影響。攜帶螢光磁性奈米鑽石的細胞可藉由奈米鑽石的螢光與磁性，分別利用流式細胞儀及磁性裝置進行分離，並可利用流式細胞儀和共軛焦顯微鏡分析細胞內螢光磁性奈米鑽石的螢光強度與位置。再者我們發現攜帶螢光磁性奈米鑽石細胞的存活率及生長能力與其原始母細胞相似。此外，攜帶螢光磁性奈米鑽石細胞內的總蛋白質表現與原始母細胞相比並沒有改變。這些分離後之攜帶螢光磁性奈米鑽石的細胞株可被冷凍及儲存，經過再解凍後的攜帶螢光磁性奈米鑽石細胞，仍具有細胞存活與生長的能力。綜合以上結果，我們建立一種新型帶有螢光磁性奈米鑽石的細胞株，將可作為生物的應用包括細胞的標定與追蹤。

Nanodiamond (ND), a carbon-based nanomaterial, has been intensively developed for biomedical applications in recent years. In this study, we created a novel ND-bearing cell line by using fluorescent and magnetic ND (FMND) after cellular separation and characterization. Treatment with FMNDs did not induce cytotoxicity in both HFL-1 normal lung fibroblasts and A549 lung cancer cells. The fluorescence intensity and particle complexity in A549 cells were elevated following exposure to FMND in a time- and concentration-dependent manner; however, the cell size distribution was not altered by FMND. FMND-bearing cells can be separated by fluorescent and magnetic properties of NDs using flow cytometer and magnetic device, respectively. The existence and fluorescence intensity of FMND in the cells were determined by flow cytometer and confocal microscope. The cell morphology, viability and growth ability of FMND-bearing cell were similar to parental cells. Moreover, the total protein expression profile of the FMND-bearing cells is not changed by comparing with the parental cells. The separated FMND-bearing cell lines can be refrigerated and stored in liquid nitrogen. After re-thawing FMND-bearing cells, the cells reserve the cell viability and growth ability. Together, we develop a novel FMND-bearing cell line, which can be provided for biological applications, including cellular labeling and tracking.