Ordered micropattern arrays fabricated by lung-derived dECM hydrogels for chemotherapeutic drug screening

Zhu, Xinglong; Li, Yi; Yang, Ying; He, Yuting; Gao, Mengyu; Peng, Wanliu; Wu, Qiong; Zhang, Guangyue; Zhou, Yanyan; Chen, Fei; Bao, Ji; Li, Wei Min

doi:10.1016/j.mtbio.2022.100274

Cited by 11 publications

(5 citation statements)

References 42 publications

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“…Furthermore, based on our previously reported study, cells cultured on micropattern arrays can exhibit matrix and homogenized characteristics because the range of cell proliferation is limited to micropatterns, allowing cells to be arranged in the matrix to form microarray observation patterns. Such as the application of micropattern arrays to high-throughput drug screening, [13][14][15] Monoclonal cell harvesting can also be done by micropattern arrays, which require varying the diameter and interpattern spacing of the micropatterns to find the right specifications. [16] Microcontact printing is one of the most convenient and time-saving methods to prepare micropatterned chips.…”

Section: Fluorescence Visualization Signals Have Been Applied In Manymentioning

confidence: 99%

Visual screening of CRISPR/Cas9 editing efficiency based on micropattern arrays for editing porcine cells

Peng,

Gao,

Zhu

et al. 2024

Biotechnology Journal

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CRISPR/Cas9 technology, combined with somatic cell nuclear transplantation (SCNT), represents the primary approach to generating gene‐edited pigs. The inefficiency in acquiring gene‐edited nuclear donors is attributed to low editing and delivery efficiency, both closely linked to the selection of CRISPR/Cas9 forms. However, there is currently no direct method to evaluate the efficiency of CRISPR/Cas9 editing in porcine genomes. A platform based on fluorescence reporting signals and micropattern arrays was developed in this study, to visually assess the efficiency of gene editing. The optimal specifications for culturing porcine cells, determined by the quantity and state of cells grown on micropattern arrays, were a diameter of 200 µm and a spacing of 150 µm. By visualizing the area of fluorescence loss and measuring the gray value of the micropattern arrays, it was quickly determined that the mRNA form targeting porcine cells exhibited the highest editing efficiency compared to DNA and Ribonucleoprotein (RNP) forms of CRISPR/Cas9. Subsequently, four homozygotes of the β4GalNT2 gene knockout were successfully obtained through the mRNA form, laying the groundwork for the subsequent generation of gene‐edited pigs. This platform facilitates a quick, simple, and effective evaluation of gene knockout efficiency. Additionally, it holds significant potential for swiftly testing novel gene editing tools, assessing delivery methods, and tailoring evaluation platforms for various cell types.

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Section: Fluorescence Visualization Signals Have Been Applied In Manymentioning

confidence: 99%

Visual screening of CRISPR/Cas9 editing efficiency based on micropattern arrays for editing porcine cells

Peng,

Gao,

Zhu

et al. 2024

Biotechnology Journal

Self Cite

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“…Using lung ECM sourced from diseased human lungs, for the generation of the hydrogels, would inherently convey (most of) the biochemical composition and resemble the mechanical properties; however, the native architecture of the lung ECM assembly is lost during the process of preparing ECM-derived hydrogels. Recently, preparing porcine lung ECM hydrogels with micropatterned surfaces was described as a method to prepare arrays for drug screening ( Zhu et al, 2022 ). This study demonstrates the preparation of spherical patterns on the hydrogel surface with different diameters, although the aspects of altering the hydrogel surface to guide cell fate, behaviour or differentiation remain unexplored.…”

Section: The Possibilities With Lung Ecm-derived Hydrogelsmentioning

confidence: 99%

Current possibilities and future opportunities provided by three-dimensional lung ECM-derived hydrogels

Nizamoglu

Burgess

2023

Front. Pharmacol.

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Disruption of the complex interplay between cells and extracellular matrix (ECM), the scaffold that provides support, biochemical and biomechanical cues, is emerging as a key element underlying lung diseases. We readily acknowledge that the lung is a flexible, relatively soft tissue that is three dimensional (3D) in structure, hence a need exists to develop in vitro model systems that reflect these properties. Lung ECM-derived hydrogels have recently emerged as a model system that mimics native lung physiology; they contain most of the plethora of biochemical components in native lung, as well as reflecting the biomechanics of native tissue. Research investigating the contribution of cell:matrix interactions to acute and chronic lung diseases has begun adopting these models but has yet to harness their full potential. This perspective article provides insight about the latest advances in the development, modification, characterization and utilization of lung ECM-derived hydrogels. We highlight some opportunities for expanding research incorporating lung ECM-derived hydrogels and potential improvements for the current approaches. Expanding the capabilities of investigations using lung ECM-derived hydrogels is positioned at a cross roads of disciplines, the path to new and innovative strategies for unravelling disease underlying mechanisms will benefit greatly from interdisciplinary approaches. While challenges need to be addressed before the maximum potential can be unlocked, with the rapid pace at which this field is evolving, we are close to a future where faster, more efficient and safer drug development targeting the disrupted 3D microenvironment is possible using lung ECM-derived hydrogels.

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“…This approach offers precise control over hydrogel architecture, which is useful for organizing cells in precise configurations and investigating how topography affects cell behavior. [103][104][105][106][107][108][109] Micromolded hydrogels, in particular, have been valuable for enhancing image-based analysis of PDOs by constraining cells to a single focal plane and reducing the artefacts resulting from imaging of cells in a 3D gel dome. For example, Brandenberg et al used micromolded PEG-based hydrogel films to create microwells that contained individual CRC PDOs restricted to a single focal plane.…”

Section: Hydrogel-based Modelsmentioning

confidence: 99%

Exploring New Dimensions of Tumor Heterogeneity: The Application of Single Cell Analysis to Organoid‐Based 3D In Vitro Models

Landon‐Brace,

Li,

McGuigan

2023

Adv Healthcare Materials

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Modelling the heterogeneity of the tumor microenvironment (TME) in vitro is essential to investigating fundamental cancer biology and developing novel treatment strategies that holistically address the factors affecting tumor progression and therapeutic response. Thus, the development of new tools for both in vitro modelling, such as patient‐derived organoids (PDOs) and complex 3D in vitro models, and single cell omics analysis, such as single‐cell RNA‐sequencing, represents a new frontier for investigating tumor heterogeneity. Specifically, the integration of PDO‐based 3D in vitro models and single cell analysis offers a unique opportunity to explore the intersecting effects of interpatient, microenvironmental, and tumor cell heterogeneity on cell phenotypes in the TME. In this review, we discuss the current use of PDOs in complex 3D in vitro models of the TME and highlight emerging directions in the development of these models. Next, we examine work that has successfully applied single cell analysis to PDO‐based models and identify important experimental considerations for this approach. Finally, we highlight open questions that may be amenable to exploration using the integration of PDO‐based models and single cell analysis. Ultimately, such investigations may facilitate the identification of novel therapeutic targets for cancer that address the significant influence of tumor‐TME interactions.This article is protected by copyright. All rights reserved

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Ordered micropattern arrays fabricated by lung-derived dECM hydrogels for chemotherapeutic drug screening

Cited by 11 publications

References 42 publications

Visual screening of CRISPR/Cas9 editing efficiency based on micropattern arrays for editing porcine cells

Visual screening of CRISPR/Cas9 editing efficiency based on micropattern arrays for editing porcine cells

Current possibilities and future opportunities provided by three-dimensional lung ECM-derived hydrogels

Exploring New Dimensions of Tumor Heterogeneity: The Application of Single Cell Analysis to Organoid‐Based 3D In Vitro Models

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