Growth of bone marrow stromal cells on small intestinal submucosa: an alternative cell source for tissue engineered bladder
Growth of bone marrow stromal cells on small intestinal submucosa: an alternative cell source for tissue engineered bladder
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DOI：
10.1111/j.1464-410x.2005.05741.x
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发表时间：
2005-11-01
影响因子：
4.5
通讯作者：
Kropp, BP
中科院分区：
文献类型：
--
作者：
Zhang, YY;Lin, HK;Kropp, BP
通过表征 BMSC 并通过免疫组织化学、生长能力和收缩性将其与膀胱平滑肌细胞 (SMC) 进行比较，评估骨髓基质细胞 (BMSC) 接种的可生物降解支架在犬模型膀胱再生中的潜在用途。通过直接针吸从五只比格犬的股骨中抽取骨髓用于体外研究。通过Ficoll-Paque (R)密度梯度离心分离单核细胞并在含有10%胎牛血清的培养基199中培养。与来自同一狗的膀胱 SMC 培养物相比，BMSC 的特征包括细胞增殖、体外收缩性、免疫组织化学分析以及小肠粘膜下层 (SIS) 支架上的生长模式。另外六只狗接受了半囊切除术，并使用接种 BMSC 的膀胱细胞（两只）、膀胱细胞（包括尿路上皮细胞加上接种 SMC 的 SIS（两只）和未接种的 SIS 支架（两只））进行膀胱扩张。增强后对六只狗进行10周的随访。体外BMSC对钙离子载体有显着的收缩反应，平均（SEM）36（2）％，相对收缩（P <0.01），这与膀胱SMC相似，但与成纤维细胞明显不同。通过免疫组织化学染色和蛋白质印迹，BMSCs还表达α-平滑肌肌动蛋白，但不表达结蛋白或肌球蛋白。在体内，接种 BMSC 和接种膀胱细胞的 SIS 移植物在整个移植物中均形成坚实的平滑肌束。BMSC 具有与原代培养的膀胱 SMC 相似的细胞增殖、组织学外观和收缩表型。 SIS 在体外支持 BMSC 的三维生长，并且接种 BMSC 的 SIS 支架促进犬模型中的膀胱再生。 BMSCs 可以作为泌尿组织工程的替代细胞来源。
To assess the potential use of bone marrow stromal cell (BMSC)-seeded biodegradable scaffold for bladder regeneration in a canine model, by characterizing BMSCs and comparing them to bladder smooth muscle cells (SMCs) by immunohistochemistry, growth capability, and contractility.Bone marrow was taken by direct needle aspiration from the femurs of five beagle dogs for the in vitro study. Mononuclear cells were isolated by Ficoll-Paque (R) density gradient centrifugation and cultivated in medium 199 with 10% fetal bovine serum. BMSCs were characterized by cell proliferation, in vitro contractility, immunohistochemical analysis, and the growth pattern on small intestinal submucosa (SIS) scaffolds compared to bladder SMC cultures from the same dogs. Another six dogs had a hemicystectomy and bladder augmentation with BMSC-seeded (two), bladder cells including urothelial cells plus SMC-seeded SIS (two) and unseeded SIS scaffolds (two). The six dogs were followed for 10 weeks after augmentation.In vitro BMSCs had a significant contractile response to calcium-ionophore, with a mean (SEM) 36 (2)%, relative contraction (P < 0.01), which was similar to bladder SMCs but markedly different from fibroblasts. BMSCs also expressed alpha-smooth muscle actin by immunohistochemical staining and Western blotting, but did not express desmin or myosin. In vivo, both BMSC-seeded and bladder cell-seeded SIS grafts had solid smooth-muscle bundle formation throughout the graft.BMSCs had a similar cell proliferation, histological appearance and contractile phenotype as primary cultured bladder SMCs. SIS supported three-dimensional growth of BMSCs in vitro, and BMSC-seeded SIS scaffold promoted bladder regeneration in a canine model. BMSCs may serve as an alternative cell source in urological tissue engineering.