Gabouev A I, Schultheiss D, Mertsching H, Köppe M, Schlote N, Wefer J, Jonas U, Stief C G
Department of Urology, Leibniz Research Laboratories for Biotechnology and Artificial Organs, Hannover Medical School, Hannover, Germany.
Int J Artif Organs. 2003 Oct;26(10):935-42. doi: 10.1177/039139880302601011.
Partial or radical cystectomy requires replacement of the urinary reservoir normally achieved by using small or large bowel segments. Our aim was to establish tissue engineering of an bioartificial bladder wall using primary cultures of porcine urothelial (pUC) and bladder smooth muscle cells (pSMC) to be reseeded on different acellular biological matrices.
Primary porcine cultures of pUC and pSMC were established from open bladder biopsy material 25 mm2 in size. Acellular matrix was generated either from a) porcine bladder wall segments or b) tubular small intestinal submucosa with the still attached decellularized muscularis layer. Reseeding of these matrices with primary cells was done in a two-dimensional static model and in a three-dimensional rotating bioreactor perfused with cell culture medium for a period of 6 weeks.
Prior to reseeding the cultured cells were characterized as pUC and pSMC by immunohistochemical staining with either anti-keratin 7 or anti-alpha actin. For both matrices a reseeded double layer cell system of pUC and pSMC could be identified after incubation in the described systems for 6 weeks.
Our results document successful generation of tissue engineered urinary bladder wall, which can be used in further large animal transplantation experiments.
部分或根治性膀胱切除术需要重建储尿囊，通常采用小肠或大肠段来实现。我们的目的是利用猪尿路上皮细胞（pUC）和膀胱平滑肌细胞（pSMC）的原代培养物，在不同的脱细胞生物基质上重新接种，建立生物人工膀胱壁的组织工程。
从大小为25平方毫米的开放膀胱活检材料中建立pUC和pSMC的猪原代培养物。脱细胞基质由以下方法制备：a）猪膀胱壁段；b）带有仍附着的脱细胞肌层的管状小肠黏膜下层。在二维静态模型和三维旋转生物反应器中，用细胞培养基灌注6周，将这些基质用原代细胞重新接种。
在重新接种之前，通过抗角蛋白7或抗α肌动蛋白的免疫组织化学染色将培养的细胞鉴定为pUC和pSMC。在所述系统中孵育6周后，两种基质都能鉴定出由pUC和pSMC重新接种的双层细胞系统。
我们的结果证明成功构建了组织工程化膀胱壁，可用于进一步的大型动物移植实验。
