螢光金奈米團簇對小鼠早期胚胎發育的影響

螢光金奈米團簇（Gold Nanoclusters , AuNCs）主要是用金奈米粒子與金原液讓粒子間相互碰撞蝕刻而成，本研究中所使用到的金奈米團簇表面嫁接上二氫硫辛酸（Dihydrolipoic acid , DHLA），成為單分子保護膜，以確保金奈米團簇不會聚集，而使得螢光金奈米團簇會有紅螢光的產生，其粒徑大小為1 - 2 nm左右；在先前的研究裡，發現螢光金奈米團簇與小鼠囊胚期（Blastocysts）的胚胎共培養的情況下，並無毒性的傷害，因此本研究想藉由螢光金奈米團簇之良好的高生物相容性與抗氧化功能，對於小鼠早期胚胎之二細胞期（two cell stage）以及桑葚胚期（Morula）胚胎與螢光金奈米團簇共培養下之探討。　　將二細胞期胚胎與螢光金奈米團簇（0 - 20 nM）共培養48小時，觀察各個處理組別之螢光金奈米團簇的螢光強度。研究結果顯示出，二細胞期胚胎在與螢光金奈米團簇10與20 nM共培養下無法順利發育到囊胚期；細胞增生實驗裡，處理組的組別與控制組相比細胞增生數均有顯著差異，凋亡分析下處理組與控制組均沒有看到凋亡訊號產生；使用共軛焦顯微鏡觀察螢光金奈米團簇的螢光分佈多停留在胚胎透明帶上，並也觀察到此材料會延滯細胞生長週期，顯示出在一定濃度範圍下螢光金奈米團簇會使得細胞之生長週期停滯，將細胞停留在單一細胞週期。

關鍵字

細胞增生；二細胞期；細胞凋亡；螢光金奈米團簇；活性氧化物質

並列摘要

Gold Nanoclusters（AuNCs）is synthesized by using a gold stock solution etched with gold nanoparticles into small nanoclusters. In this study, AuNCs were bioconjugated with dihydrolipoic acid（DHLA）for protection the AuNCs assemble, the particle size is around 1 - 2 nm which make the AuNCs to have the red fluorescent. Previous study has demonstrated that preincubation of AuNCs with mouse blastocysts has no cytotoxicity. Here, we want to further investigate the effects of AuNCs on development of mouse two cell and morula stage of embryos. Two-cell stage embryos incubated with AuNCs（0 - 20 nM）for 48 hr, with different concentrations of embryos was found to have red fluorescence, it showed that AuNCs can localized into the embryos. In this result, it showed that the embryos incubated with 10 and 20 nM of AuNCs cannot develop to blastocyst after 48 hr. In the Hoechst staining assay, we found that the total cell numbers of embryos for incubation with various concentrations of AuNCs have the significant different with control group. it shows that AuNCs have the hazardous effects on the mouse early stages embryos. In the future, we will study the effects of AuNCs on intracellular ROS changes and various stages embryo development processes in early stage embryos and mouse embryonic stem cells.

並列關鍵字

Gold nanoclusters ； reactive oxygen species ； cell apoptosis ； hoechst stain ； two cell stage

參考文獻

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螢光金奈米團簇對小鼠早期胚胎發育的影響

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