一种新颖且具有成本效益的方法,用于高通量3D培养和使用反向反射Janus微粒对hipsc衍生的心肌细胞进行节律性评估。

IF 11.3 1区 医学 Q1 Medicine
Huyen T M Pham, Duc Long Nguyen, Hyo-Sop Kim, Eun Kyeong Yang, Jae-Ho Kim, Hyun C Yoon, Hyun-Ji Park
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引用次数: 0

摘要

背景:人诱导多能干细胞来源的心肌细胞(hiPSC-CMs)作为一种有效的细胞来源在再生医学和药物发现领域受到关注。鉴于实验模型的需要,我们在此研究了一种3D培养平台和一种评估hiPSC-CMs心律的方法。方法:PAMCELL™3D细胞培养板设计用于使细胞完全附着在粘附图案区域。这些细胞粘附区被称为微图案垫,其特征是微米硅珠,表面用众所周知的精氨酸-甘氨酸-天冬氨酸(RGD)肽进行修饰。RGD与hiPSC-CMs表面的结合促进了细胞-细胞的附着和形成大小均匀的球体,这由微图案垫的直径控制。利用反向反射Janus微粒子(RJP)的反射特性对三维hiPSC-CMs加热模式进行了评估和评价。这些RJPs被一种靶向在hiPSC-CM球体表面发现的间隙连接蛋白的抗体修饰。该信号评估系统包括连接在光学显微镜上的摄像机和白光光源。结果:3D PAMCELL™R100培养板在96孔板的每孔中有效地产生约350个均匀大小的hiPSC-CM球体,并支持20天的培养。基因和蛋白表达水平分析显示,在PAMCELL™R100上生长的iPSC-CM球体保留了心脏干细胞的特征和功能,优于传统的2D培养平台。此外,RJPs可以监测和评估心肌细胞的体外跳动特性,而无需使用复杂的监测装置。当暴露于离子通道阻滞剂硝苯地平和E4031时,该系统证明了其识别心脏细胞节律活动改变的能力。结论:本研究将3D培养方法与RJPs相结合,建立了一个评估3D hiPSC-CMs节律特性的平台。这种方法在识别心律失常或其他心脏异常方面具有重要的潜力,最终有助于开发更有效的心脏病治疗方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

A novel and cost-effective method for high-throughput 3D culturing and rhythmic assessment of hiPSC-derived cardiomyocytes using retroreflective Janus microparticles.

A novel and cost-effective method for high-throughput 3D culturing and rhythmic assessment of hiPSC-derived cardiomyocytes using retroreflective Janus microparticles.

A novel and cost-effective method for high-throughput 3D culturing and rhythmic assessment of hiPSC-derived cardiomyocytes using retroreflective Janus microparticles.

A novel and cost-effective method for high-throughput 3D culturing and rhythmic assessment of hiPSC-derived cardiomyocytes using retroreflective Janus microparticles.

Background: Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) gain attention as a potent cell source in regenerative medicine and drug discovery. With the necessity of the demands for experimental models to create a more physiologically relevant model of the heart in vitro we herein investigate a 3D culturing platform and a method for assessing rhythm in hiPSC-CMs.

Methods: The 3D cell culture PAMCELL™ plate is designed to enable cells to attach exclusively to adhesive patterned areas. These cell adhesive zones, named as micro-patterned pads, feature micron silica beads that are surface-modified with the well-known arginyl-glycyl-aspartic acid (RGD) peptide. RGD binding to the surface of hiPSC-CMs facilitates cell-cell attachment and the formation of uniform-size spheroids, which is controlled by the diameter of the micro-patterned pads. The assessment and evaluation of 3D hiPSC-CMs beating pattern are carried out using reflective properties of retroreflective Janus micro-particle (RJP). These RJPs are modified with an antibody targeting the gap junction protein found on the surface of hiPSC-CM spheroids. The signal assessment system comprises a camera attached to an optical microscope and a white light source.

Results: The 3D PAMCELL™ R100 culture plate efficiently generate approximately 350 uniform-sized hiPSC-CM spheroids in each well of a 96-well plate and supported a 20-day culture. Analysis of genes and protein expression levels reveal that iPSC-CM spheroids grown on PAMCELL™ R100 retain cardiac stem cell characteristics and functions, outperforming traditional 2D culture platform. Additionally, the RJPs enable monitoring and evaluation of in vitro beating properties of cardiomyocytes without using complex monitoring setup. The system demonstrates its capability to identify alteration in the rhythmic activity of cardiac cells when exposed to ion channel blockers, nifedipine and E4031.

Conclusions: The integration of the 3D culture method and RJPs in this study establishes a platform for evaluating the rhythmic properties of 3D hiPSC-CMs. This approach holds significant potential for identifying arrhythmias or other cardiac abnormalities, ultimately contributing to the development of more effective therapies for heart diseases.

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来源期刊
Biomaterials Research
Biomaterials Research Medicine-Medicine (miscellaneous)
CiteScore
10.20
自引率
3.50%
发文量
63
审稿时长
30 days
期刊介绍: Biomaterials Research, the official journal of the Korean Society for Biomaterials, is an open-access interdisciplinary publication that focuses on all aspects of biomaterials research. The journal covers a wide range of topics including novel biomaterials, advanced techniques for biomaterial synthesis and fabrication, and their application in biomedical fields. Specific areas of interest include functional biomaterials, drug and gene delivery systems, tissue engineering, nanomedicine, nano/micro-biotechnology, bio-imaging, regenerative medicine, medical devices, 3D printing, and stem cell research. By exploring these research areas, Biomaterials Research aims to provide valuable insights and promote advancements in the biomaterials field.
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