A high throughput cell stretch device for investigating mechanobiology in vitro.

IF 6.6 3区 医学 Q1 ENGINEERING, BIOMEDICAL
APL Bioengineering Pub Date : 2024-06-26 eCollection Date: 2024-06-01 DOI:10.1063/5.0206852
Stephen J P Pratt, Christopher M Plunkett, Guray Kuzu, Ton Trinh, Joshua Barbara, Paula Choconta, Doug Quackenbush, Truc Huynh, Anders Smith, S Whitney Barnes, Joel New, James Pierce, John R Walker, James Mainquist, Frederick J King, Jimmy Elliott, Scott Hammack, Rebekah S Decker
{"title":"A high throughput cell stretch device for investigating mechanobiology <i>in vitro</i>.","authors":"Stephen J P Pratt, Christopher M Plunkett, Guray Kuzu, Ton Trinh, Joshua Barbara, Paula Choconta, Doug Quackenbush, Truc Huynh, Anders Smith, S Whitney Barnes, Joel New, James Pierce, John R Walker, James Mainquist, Frederick J King, Jimmy Elliott, Scott Hammack, Rebekah S Decker","doi":"10.1063/5.0206852","DOIUrl":null,"url":null,"abstract":"<p><p>Mechanobiology is a rapidly advancing field, with growing evidence that mechanical signaling plays key roles in health and disease. To accelerate mechanobiology-based drug discovery, novel <i>in vitro</i> systems are needed that enable mechanical perturbation of cells in a format amenable to high throughput screening. Here, both a mechanical stretch device and 192-well silicone flexible linear stretch plate were designed and fabricated to meet high throughput technology needs for cell stretch-based applications. To demonstrate the utility of the stretch plate in automation and screening, cell dispensing, liquid handling, high content imaging, and high throughput sequencing platforms were employed. Using this system, an assay was developed as a biological validation and proof-of-concept readout for screening. A mechano-transcriptional stretch response was characterized using focused gene expression profiling measured by RNA-mediated oligonucleotide Annealing, Selection, and Ligation with Next-Gen sequencing. Using articular chondrocytes, a gene expression signature containing stretch responsive genes relevant to cartilage homeostasis and disease was identified. The possibility for integration of other stretch sensitive cell types (e.g., cardiovascular, airway, bladder, gut, and musculoskeletal), in combination with alternative phenotypic readouts (e.g., protein expression, proliferation, or spatial alignment), broadens the scope of high throughput stretch and allows for wider adoption by the research community. This high throughput mechanical stress device fills an unmet need in phenotypic screening technology to support drug discovery in mechanobiology-based disease areas.</p>","PeriodicalId":46288,"journal":{"name":"APL Bioengineering","volume":null,"pages":null},"PeriodicalIF":6.6000,"publicationDate":"2024-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11210978/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"APL Bioengineering","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1063/5.0206852","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2024/6/1 0:00:00","PubModel":"eCollection","JCR":"Q1","JCRName":"ENGINEERING, BIOMEDICAL","Score":null,"Total":0}
引用次数: 0

Abstract

Mechanobiology is a rapidly advancing field, with growing evidence that mechanical signaling plays key roles in health and disease. To accelerate mechanobiology-based drug discovery, novel in vitro systems are needed that enable mechanical perturbation of cells in a format amenable to high throughput screening. Here, both a mechanical stretch device and 192-well silicone flexible linear stretch plate were designed and fabricated to meet high throughput technology needs for cell stretch-based applications. To demonstrate the utility of the stretch plate in automation and screening, cell dispensing, liquid handling, high content imaging, and high throughput sequencing platforms were employed. Using this system, an assay was developed as a biological validation and proof-of-concept readout for screening. A mechano-transcriptional stretch response was characterized using focused gene expression profiling measured by RNA-mediated oligonucleotide Annealing, Selection, and Ligation with Next-Gen sequencing. Using articular chondrocytes, a gene expression signature containing stretch responsive genes relevant to cartilage homeostasis and disease was identified. The possibility for integration of other stretch sensitive cell types (e.g., cardiovascular, airway, bladder, gut, and musculoskeletal), in combination with alternative phenotypic readouts (e.g., protein expression, proliferation, or spatial alignment), broadens the scope of high throughput stretch and allows for wider adoption by the research community. This high throughput mechanical stress device fills an unmet need in phenotypic screening technology to support drug discovery in mechanobiology-based disease areas.

用于研究体外机械生物学的高通量细胞拉伸装置。
机械生物学是一个发展迅速的领域,越来越多的证据表明,机械信号在健康和疾病中发挥着关键作用。为加速基于机械生物学的药物发现,需要新型体外系统,以适合高通量筛选的形式对细胞进行机械扰动。在此,我们设计并制造了一种机械拉伸装置和 192 孔硅胶柔性线性拉伸板,以满足基于细胞拉伸应用的高通量技术需求。为了证明拉伸板在自动化和筛选方面的实用性,我们采用了细胞分配、液体处理、高内容成像和高通量测序平台。利用该系统开发了一种检测方法,作为筛选的生物验证和概念验证读数。利用 RNA 介导的寡核苷酸退火、选择和连接以及 Next-Gen 测序技术,对基因表达进行了集中分析,从而确定了机械转录拉伸反应的特征。利用关节软骨细胞,确定了包含与软骨稳态和疾病相关的拉伸反应基因的基因表达特征。结合其他表型读数(如蛋白质表达、增殖或空间排列),可以整合其他拉伸敏感细胞类型(如心血管、气道、膀胱、肠道和肌肉骨骼),从而拓宽了高通量拉伸的范围,使研究界可以更广泛地采用。这种高通量机械应力装置满足了表型筛选技术方面尚未得到满足的需求,为基于机械生物学的疾病领域的药物发现提供了支持。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
APL Bioengineering
APL Bioengineering ENGINEERING, BIOMEDICAL-
CiteScore
9.30
自引率
6.70%
发文量
39
审稿时长
19 weeks
期刊介绍: APL Bioengineering is devoted to research at the intersection of biology, physics, and engineering. The journal publishes high-impact manuscripts specific to the understanding and advancement of physics and engineering of biological systems. APL Bioengineering is the new home for the bioengineering and biomedical research communities. APL Bioengineering publishes original research articles, reviews, and perspectives. Topical coverage includes: -Biofabrication and Bioprinting -Biomedical Materials, Sensors, and Imaging -Engineered Living Systems -Cell and Tissue Engineering -Regenerative Medicine -Molecular, Cell, and Tissue Biomechanics -Systems Biology and Computational Biology
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信