Fetal cartilage engineering from amniotic mesenchymal progenitor cells.
Fetal cartilage engineering from amniotic mesenchymal progenitor cells.
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DOI：
10.1089/scd.2006.15.245
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发表时间：
2006-04
影响因子：
4
通讯作者：
S. Kunisaki;R. Jennings;D. Fauza
中科院分区：
文献类型：
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作者：
S. Kunisaki;R. Jennings;D. Fauza
我们确定了软骨是否可以从通常在羊水中发现的间充质祖细胞（MPCs）工程。从绵羊羊水样品（n = 5）中分离间充质巨噬细胞，并通过免疫细胞化学确认其身份。扩增细胞，然后在含有转化生长因子-β 2（TGF-β 2）和胰岛素生长因子-1（IGF-1）的软骨形成培养基中以微团团块（n = 5）培养6-12周。在相同条件下培养源自胎儿真皮成纤维细胞的沉淀（n = 4）。此外，将扩增的间充质干细胞接种到可生物降解的聚乙醇酸支架上（n = 5），并在旋转生物反应器内的相同软骨形成培养基中维持10-15周。对工程化标本进行定量分析，并与天然胎儿透明软骨样品（n = 5）进行比较。统计分析通过非配对Student t检验（p < 0.05）进行。分离的细胞对波形蛋白和细胞角蛋白-8和-18染色阳性，但对CD 31染色阴性。来自间充质干细胞的微粒团通过标准和基质特异性染色表现出软骨形成分化。相比之下，这些发现不能在真皮成纤维细胞为基础的颗粒复制。来自间充质干细胞的工程化构建体同样显示出软骨形成分化的组织学证据，并保持其原始大小和三维结构。定量分析的工程结构显示较低浓度的II型胶原蛋白，但糖胺聚糖，弹性蛋白，和DNA的量相似，相比，天然胎儿透明软骨。我们的结论是，间充质干细胞可用于软骨组织工程在体外。从羊水中制造软骨可能成为外科治疗先天性畸形的一种实用方法。
We determined whether cartilage could be engineered from mesenchymal progenitor cells (MPCs) normally found in amniotic fluid. Mesenchymal amniocytes were isolated from ovine amniotic fluid samples (n = 5) and had their identity confirmed by immunocytochemistry. Cells were expanded and then cultured as micromass pellets (n = 5) in a chondrogenic medium containing transforming growth factor-beta2 (TGF-beta2) and insulin growth factor-1 (IGF-1) for 6-12 weeks. Pellets derived from fetal dermal fibroblasts (n = 4) were cultured under identical conditions. Additionally, expanded mesenchymal amniocytes were seeded onto biodegradable polyglycolic acid scaffolds (n = 5) and maintained in the same chondrogenic medium within a rotating bioreactor for 10-15 weeks. Engineered specimens were analyzed quantitatively and compared with native fetal hyaline cartilage samples (n = 5). Statistical analysis was by the unpaired Student's t-test (p < 0.05). The isolated cells stained positively for vimentin and cytokeratins-8 and -18, but negatively for CD31. Micromass pellets derived from mesenchymal amniocytes exhibited chondrogenic differentiation by both standard and matrix-specific staining. In contrast, these findings could not be replicated in dermal fibroblast-based pellets. The engineered constructs derived from mesenchymal amniocytes similarly displayed histological evidence of chondrogenic differentiation and maintained their original size and three-dimensional architecture. Quantitative assays of the engineered constructs revealed lower concentrations of collagen type II, but similar amounts of glycosaminoglycans, elastin, and DNA, when compared to native fetal hyaline cartilage. We conclude that mesenchymal amniocytes can be used for the engineering of cartilaginous tissue in vitro. Cartilage engineering from the amniotic fluid may become a practical approach for the surgical treatment of select congenital anomalies.