Phototransfection of mammalian cells using femtosecond laser pulses: optimization and applicability to stem cell differentiation
Phototransfection of mammalian cells using femtosecond laser pulses: optimization and applicability to stem cell differentiation
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DOI：
10.1117/1.3430733
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发表时间：
2010-07-01
影响因子：
3.5
通讯作者：
Gunn-Moore, Frank
中科院分区：
文献类型：
--
作者：
Mthunzi, Patience;Dholakia, Kishan;Gunn-Moore, Frank
最近，飞秒激光脉冲已被用于有针对性地将遗传物质引入哺乳动物细胞。这种使用紧密聚焦的激光束的快速扩展和发展的新光学技术被称为光转染。扩展先前的研究[史蒂文森等人，Opt. Express 14，7125-7133（2006）]，我们表明飞秒激光器可用于光致转染一系列不同的细胞系，并且具体地，这种新技术还可以以类似于25%的效率转染小鼠胚胎干细胞集落。值得注意的是，我们展示了利用光转染将这些细胞分化成胚外内胚层的能力。此外，我们提出了两个新的发现，旨在优化光转染方法，提高适用性：第一，细胞传代次数对转染效率的影响进行了探讨，第二，通过整个培养处理提高转染效率的能力。我们的研究结果应该鼓励更广泛地采用这种方法。(C)2010年，美国光学仪器工程师学会（Society of Photo-Optical Instrumentation Engineers）[DOI 10.1117/1.3430733]
Recently, femtosecond laser pulses have been utilized for the targeted introduction of genetic matter into mammalian cells. This rapidly expanding and developing novel optical technique using a tightly focused laser light beam is called phototransfection. Extending previous studies [Stevenson et al., Opt. Express 14, 7125-7133 (2006)], we show that femtosecond lasers can be used to phototransfect a range of different cell lines, and specifically that this novel technology can also transfect mouse embryonic stem cell colonies with similar to 25% efficiency. Notably, we show the ability of differentiating these cells into the extraembryonic endoderm using phototransfection. Furthermore, we present two new findings aimed at optimizing the phototransfection method and improving applicability: first, the influence of the cell passage number on the transfection efficiency is explored and, second, the ability to enhance the transfection efficiency via whole culture treatments. Our results should encourage wider uptake of this methodology. (C) 2010 Society of Photo-Optical Instrumentation Engineers. [DOI: 10.1117/1.3430733]