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首页> 外文期刊>Biomaterials Science >Protein-engineered scaffolds for in vitro 3D culture of primary adult intestinal organoids
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Protein-engineered scaffolds for in vitro 3D culture of primary adult intestinal organoids

机译：蛋白质工程支架，用于主要成人肠类器官的体外3D培养

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Though in vitro culture of primary intestinal organoids has gained significant momentum in recent years, little has been done to investigate the impact of microenvironmental cues provided by the encapsulating matrix on the growth and development of these fragile cultures. In this work, the impact of various in vitro culture parameters on primary adult murine organoid formation and growth are analyzed with a focus on matrix properties and geometric culture configuration. The air-liquid interface culture configuration was found to result in enhanced organoid formation relative to a traditional submerged configuration. Additionally, through use of a recombinantly engineered extracellular matrix (eECM), the effects of biochemical and biomechanical cues were independently studied. Decreasing mechanical stiffness and increasing cell adhesivity were found to increase organoid yield. Tuning of eECM properties was used to obtain organoid formation efficiency values identical to those observed in naturally harvested collagen I matrices but within a stiffer construct with improved ease of physical manipulation. Increased ability to remodel the surrounding matrix through mechanical or enzymatic means was also shown to enhance organoid formation. As the engineering and tunability of recombinant matrices is essentially limitless, continued property optimization may result in further improved matrix performance and may help to identify additional microenvironmental cues that directly impact organoid formation, development, differentiation, and functional behavior. Continued culture of primary organoids in recombinant matrices could therefore prove to be largely advantageous in the field of intestinal tissue engineering for applications in regenerative medicine and in vitro tissue mimics.
机译：尽管近年来主要肠道类器官的体外培养获得了巨大的发展势头，但几乎没有研究封装基质提供的微环境线索对这些脆弱培养物的生长和发育的影响。在这项工作中，以基质性质和几何培养构型为重点，分析了各种体外培养参数对成年鼠类器官的形成和生长的影响。相对于传统的淹没配置，发现气液界面培养配置可导致类器官的形成增强。此外，通过使用重组工程改造的细胞外基质（eECM），独立研究了生化和生物力学提示的作用。发现降低机械刚度和增加细胞粘附性可增加类器官的产量。 eECM特性的调整用于获得类器官形成效率值，该值与在自然收获的胶原蛋白I基质中观察到的值相同，但在更坚固的构造中具有改善的物理操作便利性。还显示出通过机械或酶促方式重塑周围基质的能力增强了类器官的形成。由于重组基质的工程设计和可调性本质上是无限的，因此持续的性能优化可能会导致基质性能进一步提高，并可能有助于确定直接影响类器官形成，发育，分化和功能行为的其他微环境提示。因此，在肠组织工程领域中，在再生医学和体外组织模拟物中的应用，可以证明在重组基质中继续培养主要类器官是非常有利的。

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