Microfluidic device for stem cell differentiation and localized electroporation of postmitotic neurons.
Microfluidic device for stem cell differentiation and localized electroporation of postmitotic neurons.
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DOI：
10.1039/c4lc00721b
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发表时间：
2014-12-07
期刊：
影响因子：
6.1
通讯作者：
Espinosa HD
中科院分区：
文献类型：
--
作者：
Kang W;Giraldo-Vela JP;Nathamgari SS;McGuire T;McNaughton RL;Kessler JA;Espinosa HD
为基因编辑和基因表达谱提供核酸和其他分子的新技术对推进生物学研究至关重要，这些技术可以在对细胞生长或分化的干扰最小的情况下进行。研究具有时间控制的自然状态下的细胞，对于从干细胞分化而来的贴壁细胞（如神经元）尤其重要。现有的高通量转染方法要么要求细胞处于悬浮状态，要么具有高毒性，并且每次实验只能进行一次转染。在这里，我们提出了一种微流控装置，将贴壁细胞的片上培养和局部电穿孔的转染结合起来。集成的微通道允许在设备上进行长期细胞培养和重复的时间转染。通过首先对HeLa和HT1080细胞进行电穿孔，验证了微流控装置的有效性，碘化丙啶的转染效率为95%，质粒的转染效率高达50%。通过芯片上的神经干细胞分化和绿色荧光蛋白质粒转染有丝分裂后神经元，证明了对原代细胞的应用。
New techniques to deliver of nucleic acids and other molecules for gene editing and gene expression profiling, which can be performed with minimal perturbation to cell growth or differentiation, are essential for advancing biological research. Studying cells in their natural state, with temporal control, is particularly important for primary cells that are derived by differentiation from stem cells and are adherent, e.g., neurons. Existing high-throughput transfection methods either require cells to be in suspension or are highly toxic and limited to a single transfection per experiment. Here we present a microfluidic device that couples on-chip culture of adherent cells and transfection by localized electroporation. Integrated microchannels allow long-term cell culture on the device and repeated temporal transfection. The microfluidic device was validated by first performing electroporation of HeLa and HT1080 cells, with transfection efficiencies of ~95% for propidium iodide and up to 50% for plasmids. Application to primary cells was demonstrated by on-chip differentiation of neural stem cells and transfection of postmitotic neurons with a green fluorescent protein plasmid.
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影响因子：
10.8
作者：
Kang W;Yavari F;Minary-Jolandan M;Giraldo-Vela JP;Safi A;McNaughton RL;Parpoil V;Espinosa HD
通讯作者：
Espinosa HD
影响因子：
6.1
作者：
Goluch, Edgar D.;Nam, Jwa-Min;Liu, Chang
通讯作者：
Liu, Chang
影响因子：
9.5
作者：
Qi, Suijian;Yi, Changqing;Yang, Mengsu
通讯作者：
Yang, Mengsu
影响因子：
6.1
作者：
Geng T;Lu C
通讯作者：
Lu C
影响因子：
48
作者：
Kitamura, Kazuo;Judkewitz, Benjamin;Hausser, Michael
通讯作者：
Hausser, Michael