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Updated: Jun 22, 2025

Decellularization and Recellularization of Whole Livers

Decellularization and Recellularization of Whole Livers

Published on: February 4, 2011

肝脏点击dECM水凝用于设计肝脏微环境.

Laura A Milton¹, Jordan W Davern², Luke Hipwood³

¹Faculty of Engineering, School of Mechanical, Medical and Process Engineering, Queensland University of Technology, Brisbane, Australia; Centre for Biomedical Technologies, Queensland University of Technology, Brisbane, Australia; Gelomics Pty Ltd, Brisbane, Australia.

Acta biomaterialia

|July 3, 2024

概括

我们从肝细胞外基质 (dECM) 开发了新的点击水凝,用于更好的3D细胞培养. 这些可重复的dECM水凝为药物测试提供了精确的机械控制和增强的肝细胞功能.

关键词:

三维细胞培养的3D细胞培养脱细胞化的细胞外矩阵.水凝是一种水凝.肝脏肝脏肝脏肝脏迈克尔类型的添加.

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科学领域:

生物材料科学生物材料科学
组织工程是组织工程.
细胞生物学细胞生物学

背景情况:

脱细胞化细胞外基质 (dECM) 水凝为3D细胞培养提供本地组织微环境,但由于不受控制的原体交叉连接,其复制能力较差.
现有的dECM水凝缺乏对机械性能的精确控制,这限制了它们对模拟各种生理和病理条件的实用性.
开发可复制和可调节的基于dECM的生物材料对于推进体外细胞培养模型至关重要.

研究的目的:

使用点击化学创建可复制和机械调节的肝脏dECM水凝系统.
优化脱细胞化协议,以最大限度地保持ECM和删除DNA.
评估新型dECM水凝在支持肝细胞中的细胞相容性和功能性能.

主要方法:

使用Triton X-100和氧化优化肝脏组织脱细胞化.
通过EDC/NHS与L-Cysteine的合,功能化素溶解肝脏dECM与硫醇 (dECM-SH).
通过dECM-SH和4臂PEG-maleimide之间的迈克尔类添加反应形成的水凝.
描述了水凝的机械性质 (Young's moduli 1-7 kPa) 并评估了3D培养中的肝细胞 (HepG2,HepaRG) 活力,增殖和功能.

主要成果:

在脱细胞化过程中实现了几乎完全的DNA去除,并保留了本地肝脏蛋白质.
合成了光学透明的,共交联的肝脏dECM水凝,具有可调节的机械性能.
与对照人群相比,证明了出色的细胞兼容性,支持了HepG2和HepaRG细胞生长,并显著增强了肝脏特异性功能 (代谢活性,CYP酶活性,分泌).
与传统的dECM水凝相比,点击dECM-SH水凝表现出更好的处理能力和可重复性.

结论:

通过点击化学形成的硫化肝 dECM 水凝为3D肝细胞培养提供了高度控制和可重复的平台.
这些新型水凝密切模仿本地肝脏的ECM特性,并允许精确调整机械特性.
在这些水凝中观察到的肝脏特异性细胞功能的增强代表了体外肝脏模型在药物发现和毒理学方面的重大进步.