Department of Urology 659, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
J Urol. 2013 Jul;190(1):341-9. doi: 10.1016/j.juro.2012.12.103. Epub 2013 Jan 8.
We developed an experimental ex vivo organoid bladder mucosal model that can be used for experimental research purposes to create alternatives to current animal models.
We developed an ex vivo organoid bladder mucosal model by immobilizing a type I collagen scaffold on the bottom of a Transwell® insert, creating a 2-compartment system. Mucosal biopsies from porcine bladders were placed on top of the scaffold and cultured in different mediums. We evaluated the morphological aspects of biopsy tissue. Cultured samples were assessed by scanning electron microscopy, and immunohistochemical and histochemical staining for cell identification, proliferation and morphology.
Cells remained viable in Dulbecco's modified Eagle's medium/F-12 and smooth muscle cell medium for up to 3 weeks. The mucosa retained normal morphological characteristics for up to 1 week. Cells (mostly urothelial cells) proliferated and fully covered the scaffold surface within 3 weeks.
We developed an experimental ex vivo organoid model of bladder mucosa for preclinical experimental research. This model could be used for high volume screening for pharmacology and toxicology experiments. It has the potential to replace currently used animal models.
我们开发了一种实验性的离体类器官膀胱黏膜模型,可用于实验研究目的,以替代当前的动物模型。
我们通过将 I 型胶原支架固定在 Transwell®插入物的底部,创建了一个 2 室系统,从而开发了离体类器官膀胱黏膜模型。将猪膀胱的黏膜活检置于支架顶部,并在不同的培养基中培养。我们评估了活检组织的形态学方面。通过扫描电子显微镜以及细胞鉴定、增殖和形态的免疫组织化学和组织化学染色来评估培养的样本。
细胞在 Dulbecco 改良 Eagle 培养基/F-12 和平滑肌细胞培养基中可存活长达 3 周。黏膜在 1 周内保留正常的形态特征。细胞(主要是尿路上皮细胞)在 3 周内增殖并完全覆盖支架表面。
我们开发了一种实验性的离体类器官膀胱黏膜模型,用于临床前实验研究。该模型可用于药理学和毒理学实验的大量筛选,有可能替代目前使用的动物模型。