膀胱平滑肌细胞与聚氨酯膜体外复合培养的实验研究

目的以聚氨酯材料为衬底进行膀胱平滑肌细胞的种植和培养,与在培养瓶中的膀胱平滑肌细胞进行对比,了解聚氨酯材料的组织相容性和细胞毒性。方法采用组织块培养法,在膀胱平滑肌细胞体外生长增殖的过程中用倒置显微镜观察其形态和大体表现,用酶标仪CCK8法检测细胞增殖的数量。结果透射电镜下在不同的倍率下观察聚氨酯膜,可以看到在不同的倍率下聚氨酯膜的表面是光滑的,材料性质单一。组织块培养法原代培养获取人膀胱平滑肌细胞稳定可靠,细胞形态良好。抗α-肌动蛋白免疫组织化学染色证实培养细胞为膀胱平滑肌细胞。膀胱平滑肌细胞在PU表面能黏附、生长和增殖。体外复合培养7 d后,膀胱平滑肌细胞铺满PU表面。5 d,7 d的细胞形态与1 d相似。两组5 d细胞计数差异有统计学意义(P<0.05)。结论膀胱平滑肌细胞在聚氨酯材料上的生长增殖与培养瓶中对比没有明显的差异,聚氨酯材料是具有良好组织相容性的生物工程材料。

Experimental studies on human bladder muscle cells on the polyurethane film

Objective To explore an effective method of culturing the human bladder smooth muscle cells, observe the morphological characteristics of the bladder smooth muscle cells grow in Polyurethane （PU） film and culture bottle. To compare the histocompatibility of using PU film and culture bottle to be as scaffoldin vitro. To offer an experimental basis for reconstruction of the bladder smooth muscle structured by the tissue engineering techniques.Methods The enzyme-treatment method and the explant method were respectively used to isolate and harvest the human bladder smooth muscle cells, and then a primary culture of these cells was performed. The human bladder smooth muscle cells were seeded on the Polyurethane （PU） film and culture bottle. At 1, 5 and 7 days of the culture, the specimens were taken; the bladder smooth muscle cells growing on the PU were observed by the inverted microscope and the number of the human BSMC.Results The composite of the bladder smooth muscle cells growing on the PU was used as the experimental group, and the BSMC growing on the culture bottle was used as the control group. In each group, 6 holes were chosen for the seeded bladder smooth muscle cells, and then the cells were collected at 1, 5 and 7 days for the cell counting after the enzyme treatment. The proliferation of the cells was assessed by the cell counting after the seeding on the PU and the culture bottle.ConclusionWith the enzyme-treatment method, the primarily cultured the human BSMC can produce a great amount of good and active cellsin vitro. The PU has good histocompatibility to support the human BSMC to adhere and grow. It has provided the tissue engineered bladder reconstruction.