微流控芯片 96 孔灌流培养系统用于高通量组织工程和长期全光学电生理学研究。
Microfluidics-enabled 96-well perfusion system for high-throughput tissue engineering and long-term all-optical electrophysiology.
机构信息
Department of Biomedical Engineering, The George Washington University, Washington, DC, USA.
出版信息
Lab Chip. 2020 Nov 7;20(21):4031-4042. doi: 10.1039/d0lc00615g. Epub 2020 Sep 30.
Abstract
This work demonstrates a novel high-throughput (HT) microfluidics-enabled uninterrupted perfusion system (HT-μUPS) and validates its use with chronic all-optical electrophysiology in human excitable cells. HT-μUPS consists of a soft multichannel microfluidic plate cover which could button on a commercial HT 96-well plate. Herein, we demonstrate the manufacturing process of the system and its usages in acute and chronic all-optical electrophysiological studies of human induced pluripotent stem-cell-derived cardiomyocytes (iPSC-CM) and engineered excitable (spiking HEK) cells. HT-μUPS perfusion maintained functional voltage and calcium responses in iPSC-CM and spiking HEK cells under spontaneous conditions and under optogenetic pacing. Long-term culture with HT-μUPS improved cell viability and optogenetically-tracked calcium responses in spiking HEK cells. The simplicity of this design and its compatibility with HT all-optical electrophysiology can empower cell-based assays for personalized medicine using patient-derived cells.
摘要
这项工作展示了一种新颖的高通量（HT）微流控不间断灌流系统（HT-μUPS），并通过人类可兴奋细胞的慢性全光学电生理学验证了其用途。HT-μUPS 由一个软多通道微流控板盖组成，可以扣在商业 HT96 孔板上。本文介绍了该系统的制造工艺及其在急性和慢性全光学电生理学研究中的应用，包括人诱导多能干细胞衍生的心肌细胞（iPSC-CM）和工程可兴奋（起搏 HEK）细胞。HT-μUPS 灌流在自发条件下和光遗传起搏下维持了 iPSC-CM 和起搏 HEK 细胞的功能电压和钙反应。HT-μUPS 的长期培养提高了起搏 HEK 细胞的细胞活力和光遗传追踪的钙反应。该设计的简单性及其与 HT 全光学电生理学的兼容性，可以为使用患者来源细胞的个性化医学提供基于细胞的检测。
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本文引用的文献
1
Cardiomyocyte maturation: advances in knowledge and implications for regenerative medicine.心肌细胞成熟：知识进展及其对再生医学的影响。
Nat Rev Cardiol. 2020 Jun;17(6):341-359. doi: 10.1038/s41569-019-0331-x. Epub 2020 Feb 3.
2
Use of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes in Preclinical Cancer Drug Cardiotoxicity Testing: A Scientific Statement From the American Heart Association.人诱导多能干细胞衍生心肌细胞在临床前癌症药物心脏毒性检测中的应用：美国心脏协会的科学声明。
Circ Res. 2019 Oct 25;125(10):e75-e92. doi: 10.1161/RES.0000000000000291. Epub 2019 Sep 19.
3
Human pluripotent stem cell-derived cardiomyocytes for studying energy metabolism.人多能干细胞衍生的心肌细胞用于研究能量代谢。
Biochim Biophys Acta Mol Cell Res. 2020 Mar;1867(3):118471. doi: 10.1016/j.bbamcr.2019.04.001. Epub 2019 Apr 4.
4
Multimodal on-axis platform for all-optical electrophysiology with near-infrared probes in human stem-cell-derived cardiomyocytes.用于人源干细胞衍生心肌细胞的近红外探针的全光学电生理学多模式共轴平台。
Prog Biophys Mol Biol. 2020 Aug;154:62-70. doi: 10.1016/j.pbiomolbio.2019.02.004. Epub 2019 Mar 5.
5
A 96-well microplate bioreactor platform supporting individual dual perfusion and high-throughput assessment of simple or biofabricated 3D tissue models.一种 96 孔微孔板生物反应器平台，支持单个双灌注和高通量评估简单或生物制造的 3D 组织模型。
Lab Chip. 2018 Sep 11;18(18):2757-2775. doi: 10.1039/c8lc00485d.
6
Multiparametric slice culture platform for the investigation of human cardiac tissue physiology.用于研究人类心脏组织生理学的多参数切片培养平台。
Prog Biophys Mol Biol. 2019 Jul;144:139-150. doi: 10.1016/j.pbiomolbio.2018.06.001. Epub 2018 Jun 28.
7
Soft Lithography.软光刻
Angew Chem Int Ed Engl. 1998 Mar 16;37(5):550-575. doi: 10.1002/(SICI)1521-3773(19980316)37:5<550::AID-ANIE550>3.0.CO;2-G.
8
Advanced maturation of human cardiac tissue grown from pluripotent stem cells.多能干细胞来源的人心肌组织的高级成熟。
Nature. 2018 Apr;556(7700):239-243. doi: 10.1038/s41586-018-0016-3. Epub 2018 Apr 4.
9
Silicones for Stretchable and Durable Soft Devices: Beyond Sylgard-184.用于可拉伸和耐用的软设备的硅酮：超越 Sylgard-184。
ACS Appl Mater Interfaces. 2018 Apr 4;10(13):11261-11268. doi: 10.1021/acsami.7b18394. Epub 2018 Mar 26.
10
Cardiac performance is limited by oxygen delivery to the mitochondria in the crystalloid-perfused working heart.心脏的功能受限于晶体液灌注工作心脏中线粒体的氧供。
Am J Physiol Heart Circ Physiol. 2018 Apr 1;314(4):H704-H715. doi: 10.1152/ajpheart.00321.2017. Epub 2017 Nov 10.
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