在化学定义的,无动物起源的细胞培养中重建人类表皮的表征
Julia Bajsert1,2, Valérie De Glas1, Emilie Faway1
1NARILIS-Faculty of Medicine, University of Namur, Namur, Belgium.
概括
化学定义的无动物源 (cdAOF) 补充剂成功重建人体表皮 (RHE) 模型. 这种伦理替代方法显示了与传统方法相比较的组织学和屏障功能,为先进的体外毒理学铺平了道路.
科学领域:
- 在体外毒理学和皮肤学研究.
- 再生医学和组织工程.
- 动物替代试验方法
背景情况:
- 重建的人类表皮 (RHE) 模型为皮肤病学和毒理学中动物试验提供了伦理替代方案.
- 传统的RHE模型通常依赖于含有动物衍生成分的补充剂,例如牛体垂体提取物.
- 商业上可用的化学定义的无动物原产物 (cdAOF) 补充剂是动物衍生成分的潜在替代品.
研究的目的:
- 为了比较商业上可用的cdaof补充剂与RHE模型重建的传统补充剂的疗效.
- 用各种状细胞来源 (新生儿,成年人,不朽化) 在cdaof条件下培养的RHE模型的特征.
- 评估在cDAOF介质中培养的RHE模型的组织学特征,标记物表达和屏障功能.
主要方法:
- 使用cDAOF介质从不同的角质细胞来源培养RHE模型.
- 与使用标准的人类角质细胞生长补充剂培养的RHE模型进行比较.
- 组织学分析,特定标记物的免疫定位 (例如,involutrin),mRNA表达分析,以及跨体电阻 (TEER) 测量以评估屏障功能.
主要成果:
- 在cdAOF介质中培养的RHE模型表现出与在传统介质中培养的类似的组织学特征,基底角质细胞形态略有差异.
- 在cdaof条件下观察到卷膜蛋白的免疫定位和炎症增殖标记的mRNA表达的增加.
- 通过TEER评估的屏障功能在cDAOF培养的RHE中与对照组相比,在统计上可比或略低.
结论:
- 使用cDAOF培养条件可以实现RHE模型的重建.
- cdAOF补充剂为RHE模型生成提供了一个可行的,道德的替代方案.
- 对cdaof介质的进一步改进可以将其在体外毒理学中的应用扩展到再生医学之外.
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