Fujisaki Hitomi, Adachi Eijiro, Hattori Shunji
Nippi Research Institute of Biomatrix Toride, Ibaraki, Japan.
Connect Tissue Res. 2008;49(6):426-36. doi: 10.1080/03008200802324998.
We examined the behavior of human foreskin keratinocytes (HFKs) on reconstituted type IV collagen gel. HFKs survived for several days and the upper layer cells expressed a differentiation marker, involucrin. Apoptosis was induced after involucrin expression while cell proliferation was suppressed. On molecular type IV collagen, integrins shifted from alpha 2 beta 1 to alpha 3 beta 1 during HFK culture. On type IV collagen gel, HFKs initially expressed integrin alpha 2 beta 1, and later expressed integrin alpha 3 beta 1 in the presence of alpha 2 beta 1 did not disappear. Using synthetic peptides, we examined integrin alpha2-mediated adhesion to type IV collagen gel. Addition of synthetic peptide dose-dependently inhibited cell adhesion both on type IV collagen gel and on molecular type IV collagen. On type IV collagen gel, weaker phosphorylation of focal adhesion kinase, paxillin, and Akt was observed compared with the molecular forms. Based on these observations, we think type IV collagen gel is a novel culture substrate that mimics the physiological environment for HFKs.
我们研究了人包皮角质形成细胞（HFK）在重组IV型胶原凝胶上的行为。HFK存活了数天，上层细胞表达了一种分化标志物——兜甲蛋白。兜甲蛋白表达后诱导了细胞凋亡，同时细胞增殖受到抑制。在分子IV型胶原上，整合素在HFK培养过程中从α2β1转变为α3β1。在IV型胶原凝胶上，HFK最初表达整合素α2β1，随后在α2β1存在的情况下表达整合素α3β1且未消失。我们使用合成肽研究了整合素α2介导的对IV型胶原凝胶的黏附。合成肽的添加以剂量依赖的方式抑制了细胞在IV型胶原凝胶和分子IV型胶原上的黏附。在IV型胶原凝胶上，观察到粘着斑激酶、桩蛋白和Akt的磷酸化程度较弱。基于这些观察结果，我们认为IV型胶原凝胶是一种模拟HFK生理环境的新型培养底物。
