Department of Bioengineering, University of Washington, 3720 15th Ave NE, Seattle, WA, 98105, USA.
Adv Biol (Weinh). 2022 Nov;6(11):e2200129. doi: 10.1002/adbi.202200129. Epub 2022 Jun 30.
The human colon plays a critical role in fluid and salt absorption and harbors the largest immune compartment. There is a widespread need for in vitro models of human colon physiology with its innate immune system. A method is described to produce a cassette with a network of struts supporting a suspended, non-chemically cross-linked collagen hydrogel scaffold compatible with the co-culture of primary gastrointestinal epithelium and migratory inflammatory cells. The epithelial monolayer cultured on the suspended collagen possesses a population of polarized and differentiated cells similar to that present in vivo. This epithelial layer displays proper barrier function with a transepithelial electrical resistance (TEER) ≥ 1,500 Ω cm and an apparent permeability ≤10 cm s . Immune cells plated on the basal face of the scaffold transmigrated over a period of 24 h to the epithelial layer in response to epithelial production of IL-8 induced by luminal stimulation of Clostridium difficile Toxin A. These studies demonstrate that this in vitro platform possesses a functional primary colonic epithelial layer with an immune cell compartment capable of recruitment in response to pro-inflammatory cues coming from the epithelium.
人类结肠在液体和盐吸收中起着关键作用，并且是最大的免疫部位。人们普遍需要具有固有免疫系统的体外人类结肠生理学模型。本文描述了一种制作卡匣的方法，该卡匣具有一个由支柱支撑的网络，可支撑一个悬浮的、非化学交联的胶原水凝胶支架，该支架与原代胃肠道上皮细胞和迁移性炎症细胞共培养兼容。在悬浮胶原上培养的上皮单层具有类似于体内存在的极化和分化细胞群体。该上皮层具有适当的屏障功能，跨上皮电阻 (TEER) ≥ 1,500 Ω cm，表观渗透率≤10 cm s。免疫细胞在支架的基底面上种植，并在 24 小时内迁移到上皮层，这是对上皮细胞产生的 IL-8 的反应，上皮细胞产生的 IL-8 是由腔内刺激艰难梭菌毒素 A 引起的。这些研究表明，这种体外平台具有功能性的原代结肠上皮层，以及能够对来自上皮的促炎信号做出反应的免疫细胞群。
