Yuan Haichao, Liu Liangren, Zheng Shuo, Liu Zhenhua, Yang Lu, Pu Chunxiao, Li Jinhong, Long Dan, Wei Qiang, Han Ping
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2013 Dec;27(12):1506-11.
To observe whether umbilical cord mesenchymal stem cells (UCMSCs) can differentiate into the smooth muscle cells (SMCs) induced by bladder SMCs (BSMCs) conditioned medium so as to seek an alternative seed cells for the repair and reconstruction of the urology system.
UCMSCs and BSMCs were harvested from umbilical cord of full-term births and bladder tissues which were obtained from patients who underwent a radical cystectomy. BSMCs conditioned medium was prepared by mixing supernatant of BSMCs at passages 1-5 with complete medium at ratio of 1:1. UCMSCs at passage 3 were cultured with BSMCs conditioned medium (induced group, group A) and complete medium (control group, group B), respectively; simple BSMCs served as positive control group (group C). The morphological changes of co-cultured UCMSCs were observed by inverted phase microscope, the expressions of a-smooth muscle actin (a-SMA), Calponin, and smooth muscle myosin heavy chain (SM-MHC) of UCMSCs were tested by immunofluorescence staining and Western blot at 7 and 14 days.
The morphology of UCMSCs in group A started to change from a polygonal and short spindle shape to a large and spindle shape after co-culture, which was similar to BSMCs morphology; but the morphology of UCMSCs did not change obviously in group B. Immunofluorescence staining showed that the expressions of alpha-SMA, Calponin, and SM-MHC were positive in group C. At 7 days, the expression of a-SMA could be observed in groups A and B; at 14 days, the positive expression of alpha-SMA increased gradually in group A, but it did not increase in group B. At 7 days, a positive expression of Calponin could be observed in group A, and positive expression increased obviously at 14 days; the expression of Calponin could not be observed at 7 and 14 days in group B. However, the expression of SM-MHC could not be observed in groups A and B. The results of Western blot showed the expressions of alpha-SMA, Calponin, and SM-MHC protein were consistent with the results of immunofluorescence staining.
UCMSCs have the potential of differentiation into SMCs and may be a potential seed cells for bladder tissue engineering.
观察脐带间充质干细胞（UCMSCs）能否在膀胱平滑肌细胞（BSMCs）条件培养基诱导下分化为平滑肌细胞（SMCs），以期寻找一种用于泌尿外科系统修复与重建的替代种子细胞。
从足月产脐带和接受根治性膀胱切除术患者的膀胱组织中获取UCMSCs和BSMCs。将第1 - 5代BSMCs的上清液与完全培养基按1:1比例混合制备BSMCs条件培养基。将第3代UCMSCs分别用BSMCs条件培养基培养（诱导组，A组）和完全培养基培养（对照组，B组）；单纯BSMCs作为阳性对照组（C组）。通过倒置相差显微镜观察共培养的UCMSCs的形态变化，在第7天和第14天通过免疫荧光染色和蛋白质印迹法检测UCMSCs中α - 平滑肌肌动蛋白（α - SMA）、钙调蛋白和平滑肌肌球蛋白重链（SM - MHC）的表达。
A组UCMSCs共培养后形态开始从多边形和短梭形变为大的梭形，与BSMCs形态相似；但B组UCMSCs形态无明显变化。免疫荧光染色显示C组α - SMA、钙调蛋白和SM - MHC表达呈阳性。第7天，A组和B组均可观察到α - SMA表达；第14天，A组α - SMA阳性表达逐渐增加，而B组未增加。第7天，A组可观察到钙调蛋白阳性表达，第14天阳性表达明显增加；B组在第7天和第14天均未观察到钙调蛋白表达。然而,A组和B组均未观察到SM - MHC表达。蛋白质印迹法结果显示α - SMA、钙调蛋白和SM - MHC蛋白表达与免疫荧光染色结果一致。
UCMSCs具有分化为SMCs的潜能，可能是膀胱组织工程的潜在种子细胞。
