Transfection in perfused microfluidic cell culture devices: A case study.
Transfection in perfused microfluidic cell culture devices: A case study.
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DOI：
10.1016/j.procbio.2016.09.006
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发表时间：
2017-08
期刊：
影响因子：
--
通讯作者：
Szita N
中科院分区：
文献类型：
--
作者：
Raimes W;Rubi M;Super A;Marques MPC;Veraitch F;Szita N
用于长期均质细胞培养的微流体装置中的自动化学转染。转染试剂混合物均匀暴露于在装置中生长的细胞。相对于24孔板，用GFP转染的胚胎干细胞在芯片上显示出提高的效率和增加的中值荧光强度。自动化微流体装置是实现即时自体细胞治疗的一种有前途的途径。诱导多能干细胞（iPSC）衍生的初始步骤涉及转染和长期细胞培养。这些步骤的整合将有助于降低微尺度设备的成本和占地面积，并应用于细胞重编程或基因校正。目前转染整合的例子集中在最大化效率，而不是可行的长期培养。在这里，我们通过将自动化转染与为均匀培养条件设计的灌注微流体装置相结合来寻找整个过程的兼容性。使用荧光素对进样过程进行表征，以建立基于LabVIEW的用户自定义自动化程序。概念验证通过将GFP质粒化学转染到小鼠胚胎干细胞（mESC）中来证明。与标准方案（17.2%，n = 3）相比，在装置中转染的细胞显示出效率的提高（34%，n = 3）。这代表了迈向用于细胞重编程或基因治疗的微流体处理系统的第一步。
Automated chemical transfection in a microfluidic device for long-term homogeneous cell culture. Transfection reagent mixture uniformly exposed to cells growing in the device. Embryonic stem cells transfected with GFP show improved efficiency and an increased median fluorescence intensity on-chip relative to 24 well plate. Automated microfluidic devices are a promising route towards a point-of-care autologous cell therapy. The initial steps of induced pluripotent stem cell (iPSC) derivation involve transfection and long term cell culture. Integration of these steps would help reduce the cost and footprint of micro-scale devices with applications in cell reprogramming or gene correction. Current examples of transfection integration focus on maximising efficiency rather than viable long-term culture. Here we look for whole process compatibility by integrating automated transfection with a perfused microfluidic device designed for homogeneous culture conditions. The injection process was characterised using fluorescein to establish a LabVIEW-based routine for user-defined automation. Proof-of-concept is demonstrated by chemically transfecting a GFP plasmid into mouse embryonic stem cells (mESCs). Cells transfected in the device showed an improvement in efficiency (34%, n = 3) compared with standard protocols (17.2%, n = 3). This represents a first step towards microfluidic processing systems for cell reprogramming or gene therapy.
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