Peterson Erik T K, Papautsky Ian
Department of Electrical and Computer Engineering and Computer Science, University of Cincinnati, OH, USA.
Methods Mol Biol. 2006;321:179-97. doi: 10.1385/1-59259-997-4:179.
Musculoskeletal tissue-engineering strategies have recently focused on the use of biomaterial scaffolds capable of guiding growth and organization of mesenchymal stem cells (MSCs), which are precursors for connective tissues. This chapter describes the methods for culturing MSCs on micropatterned polydimethylsiloxane (PDMS) substrates. MSCs are isolated from bone marrow biopsies and subcultivated before plating onto PDMS substrates. Micropatterned substrates are fabricated by casting PDMS on AZ P4620 photoresist molds. Prior to plating cells, substrates are cleaned, sterilized, and coated with fibronectin. Micropatterned growth surfaces are a useful research tool enabling the study of cell morphology and alignment in response to substrate geometry. Understanding MSC response to surface topography will assist in the design of improved scaffolds for connective-tissue repair.
肌肉骨骼组织工程策略最近集中于使用能够引导间充质干细胞（MSCs）生长和组织的生物材料支架，间充质干细胞是结缔组织的前体。本章描述了在微图案化聚二甲基硅氧烷（PDMS）基质上培养间充质干细胞的方法。间充质干细胞从骨髓活检中分离出来，并在接种到PDMS基质上之前进行传代培养。通过将PDMS浇铸在AZ P4620光刻胶模具上来制造微图案化基质。在接种细胞之前，对基质进行清洗、灭菌并涂覆纤连蛋白。微图案化生长表面是一种有用的研究工具，能够研究细胞形态以及细胞对基质几何形状的排列响应。了解间充质干细胞对表面形貌的反应将有助于设计用于结缔组织修复的改良支架。
