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首页> 外文会议>Conference on optical interactions with tissue and cells >Femtosecond laser assisted photo-transfection and differentiation of mouse embryonic stem cells
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Femtosecond laser assisted photo-transfection and differentiation of mouse embryonic stem cells

机译：飞秒激光辅助小鼠胚胎干细胞的光转染和分化

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摘要

In tissue engineering research, stem cells have been used as starting material in the synthesis of mammalian cells for the treatment of various cell based diseases. This is done by manipulating the DNA content of the cells to induce a specific effect such as increased proliferation or developing a new cell type through the process of differentiation. Such controlled gene expression of stem cells is achieved by the method of transfection, where exogenous plasmid deoxyribonucleic acid (pDNA) is inserted into a stem cell using chemical, viral or physical methods. In this research, we used femtosecond (fs) laser pulses from a home-build microscope system to perforate the cellular membrane and allow entry of selected pDNA to alter the behaviour of mouse embryonic stem cells (mESCs). In one set of experiments, we induce fluorescence on mESCs using green fluorescence protein plasmid (pGFP) while in other tests; differentiation of mESCs into endoderm cells is performed using Sox-17 plasmid DNA (pSox-17). Primitive endoderm formation was thereafter confirmed using polymerase chain reactions (PCR) and the Sox-17 primer. Cell viability studies using adenosine triphosphate were also conducted. From the data, it was concluded that the photo-transfection method is biocompatible since it was able to induce fluorescence in mESCs. Secondly, it was confirmed that Sox-17 was photo-transfected successfully using 6 μW laser power, 128 fs pulses and 1kHz pulse repetition rate.
机译：在组织工程研究中，干细胞已被用作合成哺乳动物细胞以治疗各种基于细胞的疾病的原料。这可以通过操纵细胞的DNA含量来诱导特定的作用来实现，例如通过分化过程增加增殖或开发新的细胞类型。干细胞的这种受控基因表达是通过转染方法实现的，其中使用化学，病毒或物理方法将外源质粒脱氧核糖核酸（pDNA）插入干细胞中。在这项研究中，我们使用了来自家用显微镜系统的飞秒（fs）激光脉冲在细胞膜上穿孔并允许选定的pDNA进入，从而改变小鼠胚胎干细胞（mESCs）的行为。在一组实验中，我们使用绿色荧光蛋白质粒（pGFP）在mESC上诱导荧光，而在其他实验中；使用Sox-17质粒DNA（pSox-17）将mESC分化为内胚层细胞。此后，使用聚合酶链反应（PCR）和Sox-17引物确认了原始内胚层的形成。还进行了使用三磷酸腺苷的细胞活力研究。从数据得出的结论是，光转染方法具有生物相容性，因为它能够在mESCs中诱导荧光。其次，证实了使用6μW激光功率，128 fs脉冲和1kHz脉冲重复频率成功地成功转染了Sox-17。

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  • 来源
  • 会议地点 San Francisco(US)
  • 作者

    Lebogang Thobakgale; Sello Manoto; Saturnin Ombinda Lemboumba; Malik Maaza; Patience Mthunzi-Kufa;

  • 作者单位

    National Laser Centre, Council for Scientific and Industrial Research, PO Box 395,Pretoria, 0001, South Africa,Department of Physics, University of South Africa (UNISA) PO Box 392, Pretoria, 0003,South Africa;

    National Laser Centre, Council for Scientific and Industrial Research, PO Box 395,Pretoria, 0001, South Africa;

    National Laser Centre, Council for Scientific and Industrial Research, PO Box 395,Pretoria, 0001, South Africa;

    Department of Physics, University of South Africa (UNISA) PO Box 392, Pretoria, 0003,South Africa;

    National Laser Centre, Council for Scientific and Industrial Research, PO Box 395,Pretoria, 0001, South Africa,Department of Physics, University of South Africa (UNISA) PO Box 392, Pretoria, 0003,South Africa;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Photo-transfection; femtosecond; differentiation pGFP; pSox-17; primitive endoderm;

    机译：光转染；飞秒分化pGFP; pSox-17；原始内胚层;

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