國立臺灣大學醫學院復健科陳文翔2006-07-262018-07-132006-07-262018-07-132004-07-31http://ntur.lib.ntu.edu.tw//handle/246246/28983如何發展安全及有效的基因傳送方法在基因治療中一直是主要的挑戰，本研 究主旨在於找出最佳化的超音波聲參數使基因轉移效率以及細胞存活率皆能增 加，我們有興趣的參數包括超音波脈衝重複頻率(pulse repetition frequency)、 暫停時間(off time)以及時間比率對體外培養細胞的影響。本研究中發現，對於子 宮頸癌細胞(HeLa cell)而言，當超音波工作週期(duty cycle)為50%時，最佳脈 衝重複頻率為20 Hz 到1000 Hz 之間；當脈衝重複頻率低於2 Hz 時，細胞擁 有高轉移效率，但會造成高死亡率及高溫升；當脈衝重複頻率高於1000 Hz 時， 轉移效率明顯開始下降，而當脈衝重複頻率高於4000 Hz 時，細胞雖然有高存 活率及低溫升，但轉移效率卻為0%。另外由實驗結果中可發現，超音波每一次 的施打時間與暫停時間比例(on-time/off-time)需大於0.1，當此比例小於0.1 時， 轉移效率急速下降。It is always a great challenge to develop a safe and effective method for delivering gene into cells. The main purpose of this study is to determine optimal acoustic parameters for effective gene transfer and high cell survival in the ultrasound-mediated gene transfection. The parameters we studied included the pulse repetition frequency (PRF), the length off time and the ratio of on to off time (on-off ratio). We found that for cultured cervical cancer cells (HeLa cells), the best PRF for gene transfection ranges between 2 and 2000 Hz (50% duty cycle). The transfection efficiency declined rapidly while PRF was higher than 1000Hz and dropped to 1% while PRF was 4000 Hz. When the ultrasound pulse repetition frequency (PRF) was shorter than 2 Hz (the exposure on-time was larger than 250-ms), the transfection efficiency was high, but the extremely high temperature elevation resulted in low cell survival. While PRF was between 20 Hz to 1000 Hz, a better transfection could be obtained when the ratio of the on-time to the off time was larger than 0.1, i.e. > 0.1 − − off time on time . When the ratio was smaller than 0.1, the transfection efficiency was close to 0%.application/pdf103353 bytesapplication/pdfzh-TW國立臺灣大學醫學院復健科超音波基因治療基因轉移ultrasoundgene therapygene transfergenetransfection[SDGs]SDG3otherhttp://ntur.lib.ntu.edu.tw/bitstream/246246/28983/1/922312B002019.pdf