IRCCS Policlinico S. Donato, via Morandi 30, 20097, S. Donato Milanese, Milano, Italy.
Biomaterials. 2011 Jan;32(1):119-27. doi: 10.1016/j.biomaterials.2010.09.003. Epub 2010 Nov 11.
A key challenge in tissue engineering a heart valve is to reproduce the major tissue structures responsible for native valve function. Here we evaluated human adipose-derived stem cells (ADSCs) as a source of cells for heart valve tissue engineering investigating their ability to synthesize and process collagen and elastin. ADSCs were compared with human bone marrow mesenchymal stem cells (BmMSCs) and human aortic valve interstitial cells (hVICs). ADSCs and BmMSCs were stretched at 14% for 3 days and collagen synthesis determined by [(3)H]-proline incorporation. Collagen and elastin crosslinking was assessed by measuring pyridinoline and desmosine respectively, using liquid chromatography/mass spectrometry. Three-dimensional culture was obtained by seeding cells onto bovine collagen type I scaffolds for 2-20 days. Expression of matrix proteins and processing enzymes was assessed by Real Time-PCR, immunofluorescence and transmission electron microscopy. Stretch increased the incorporation of [(3)H]-proline in ADSCs and BmMSCs, however only ADSCs and hVICs upregulated COL3A1 gene. ADSCs produced collagen and elastin crosslinks. ADSCs uniformly populated collagen scaffolds after 2 days, and fibrillar-like collagen was detected after 20 days. ADSCs sense mechanical stimulation and produce and process collagen and elastin. These novel findings have important implications for the use of these cells in tissue engineering.
组织工程心脏瓣膜的一个关键挑战是复制负责天然瓣膜功能的主要组织结构。在这里，我们评估了人脂肪来源干细胞（ADSCs）作为心脏瓣膜组织工程细胞来源的潜力，研究了它们合成和加工胶原蛋白和弹性蛋白的能力。将 ADSCs 与骨髓间充质干细胞（BMSCs）和人主动脉瓣间质细胞（hVICs）进行了比较。将 ADSCs 和 BMSCs 拉伸至 14%并持续 3 天，通过 [(3)H]-脯氨酸掺入来确定胶原蛋白合成。通过使用液相色谱/质谱法分别测量吡啶酚和脱甲硅氧烷来评估胶原蛋白和弹性蛋白交联。通过将细胞接种到牛胶原蛋白 I 支架上 2-20 天来获得三维培养。通过实时 PCR、免疫荧光和透射电子显微镜评估基质蛋白和加工酶的表达。拉伸增加了 ADSCs 和 BMSCs 中 [(3)H]-脯氨酸的掺入，但只有 ADSCs 和 hVICs 上调了 COL3A1 基因。ADSCs 产生胶原蛋白和弹性蛋白交联。ADSCs 在 2 天后均匀地填充胶原蛋白支架，并且在 20 天后检测到纤维状胶原蛋白。ADSCs 感知机械刺激并产生和加工胶原蛋白和弹性蛋白。这些新发现对这些细胞在组织工程中的应用具有重要意义。
