Greenleaf W J, Bolander M E, Sarkar G, Goldring M B, Greenleaf J F
Mayo High School, Rochester, MN, USA.
Ultrasound Med Biol. 1998 May;24(4):587-95. doi: 10.1016/s0301-5629(98)00003-9.
The efficiency of ultrasound-mediated gene transfection was enhanced three- to fourfold, compared to previous results, through the use of green fluorescent protein reporter gene, cultured immortalized human chondrocytes and artificial cavitation nuclei in the form of Albunex. Cells were exposed to 1.0-MHz ultrasound transmitted through the bottom of six-well culture plates containing immortalized chondrocytes, media, DNA at a concentration of 40 micrograms/mL and Albunex at 50 x 10(6) bubbles/mL. Transfection efficiency increased linearly with ultrasound exposure pressure with a transfection threshold observed at a spatial average peak positive pressure (SAPP) of 0.12 MPa and reaching about 50% of the living cells when exposed to 0.41 MPa SAPP for 20 s. Adding fresh Albunex at 50 x 10(6) bubbles/mL prior to sequential 1-s, 0.32- or 0.41-MPa exposures increased transfection with each exposure, reaching 43% transfection after four exposures. Efficient in vitro and in vivo transfection now appear possible with these enhancements.
与先前的结果相比，通过使用绿色荧光蛋白报告基因、培养的永生化人软骨细胞以及以Albunex形式存在的人工空化核，超声介导的基因转染效率提高了三到四倍。将细胞暴露于通过含有永生化软骨细胞、培养基、浓度为40微克/毫升的DNA以及浓度为50×10⁶个气泡/毫升的Albunex的六孔培养板底部传输的1.0兆赫超声中。转染效率随超声暴露压力呈线性增加，在空间平均峰值正压（SAPP）为0.12兆帕时观察到转染阈值，当暴露于0.41兆帕SAPP 20秒时，约50%的活细胞发生转染。在依次进行1秒、0.32兆帕或0.41兆帕暴露之前，添加浓度为50×10⁶个气泡/毫升的新鲜Albunex，每次暴露都会增加转染率，四次暴露后转染率达到43%。通过这些改进，现在体外和体内的高效转染似乎成为可能。
