Young Tai-Horng, Tu Hui-Ru, Chan Chih-Chieh, Huang Yi-Ching, Yen Meng-Hua, Cheng Nai-Chen, Chiu Hsien-Ching, Lin Sung-Jan
Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, No. 1, Sec. 1, Jen-Ai Road, Taipei 100, Taiwan.
Biomaterials. 2009 Oct;30(28):5031-40. doi: 10.1016/j.biomaterials.2009.05.065. Epub 2009 Jun 24.
Generally, cells tend to aggregate on a substratum with lower cell adhesivity. However, it also leads to compromised cell growth and higher cell loss after seeding. This study is aimed at tackling this dilemma by extracellular matrix (ECM) protein coating of a lower adhesive substratum poly(ethylene-co-vinyl alcohol) (EVAL) that has been shown to facilitate hair follicle dermal papilla (DP) spheroid formation. We found that coating with either fibronectin (Fn), collagen I, or collagen IV yields higher adhesivity and cell growth than that with laminin. However, cells can only aggregate on uncoated or Fn-coated EVAL. Quantitatively, Fn coating increases the number of spheroids by 67%. Analysis of cell migration reveals that collagen I, collagen IV and laminin coatings reduce cell motility, while Fn coating keeps cells highly motile. Inhibition of cell migration hinders spheroid formation. In addition, disruption of Fn function does not significantly compromise intercellular adhesion. Hence, Fn enhances cell aggregation by enhancing cell attachment, cell growth and cell motility. Our study demonstrates that intercellular organization as spheroids or flat monolayers is switchable by specific ECM protein coating and preserving cell motility is vital to cell aggregation. In addition to generation of spheroidal DP microtissues for hair follicle regeneration and large-scale production of aggregates of other cells, this strategy can help to regulate the tissue-substrate adhesivity and tissue spreadability on the surface of implantable materials.
一般来说，细胞倾向于聚集在细胞粘附性较低的基质上。然而，这也会导致接种后细胞生长受损和细胞损失增加。本研究旨在通过在低粘附性基质聚乙烯醇共聚物（EVAL）上进行细胞外基质（ECM）蛋白包被来解决这一难题，该基质已被证明有助于毛囊真皮乳头（DP）球体的形成。我们发现，用纤连蛋白（Fn）、I型胶原蛋白或IV型胶原蛋白包被比用层粘连蛋白包被具有更高的粘附性和细胞生长。然而，细胞只能在未包被或Fn包被的EVAL上聚集。定量分析表明，Fn包被使球体数量增加了67%。细胞迁移分析表明，I型胶原蛋白、IV型胶原蛋白和层粘连蛋白包被会降低细胞运动性，而Fn包被使细胞保持高运动性。抑制细胞迁移会阻碍球体形成。此外，Fn功能的破坏不会显著损害细胞间粘附。因此，Fn通过增强细胞附着、细胞生长和细胞运动性来促进细胞聚集。我们的研究表明，通过特定的ECM蛋白包被，细胞间组织可以从球体转变为扁平单层，并且保持细胞运动性对细胞聚集至关重要。除了生成用于毛囊再生的球状DP微组织以及大规模生产其他细胞的聚集体外，该策略还可以帮助调节可植入材料表面的组织 - 基质粘附性和组织铺展性。
