Division of Education for Global Standard, Institute of Liberal Arts & Science, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan.
Division of Molecular Virology & Oncology, Cancer Research Institute, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan.
Biotechniques. 2018 Sep;65(3):137-142. doi: 10.2144/btn-2018-0036.
A modified invasion assay using a three-dimensional collagen gel was developed that enables isolation of invasive living cells; it was named the invading cell trapping (iCT) assay. A small cell strainer consisting of a nylon mesh with 40-μm pores was used, and collagen gel layers formed across the membrane. Test cells were seeded in the lower gel layer and invasive cells were attracted upward and trapped in the upper gel. After incubation, the collagen gel layers in cell strainers were easily separated and living cells in the gel were counted and analyzed. An advantage of the iCT assay is that it can capture living invasive cells in the upper gel while leaving noninvasive ones in the lower layer. Further enrichment of the two cell populations can be achieved by repeating the assay. Thus, the iCT assay allows comparative analysis of invasive versus noninvasive cells.
我们开发了一种改良的侵袭分析实验，使用三维胶原凝胶来分离侵袭性活细胞，我们称之为侵袭细胞捕获（iCT）实验。该实验使用一个带有 40 微米孔的尼龙网制成的小细胞过滤器，在膜上形成胶原凝胶层。测试细胞接种在下层凝胶中，向上迁移的侵袭细胞会被吸引并困在上层凝胶中。孵育后，细胞过滤器中的胶原凝胶层很容易分离，凝胶中的活细胞可以被计数和分析。iCT 实验的一个优点是，它可以捕获上层凝胶中的活侵袭细胞，而将非侵袭细胞留在下层。通过重复实验，可以进一步富集这两种细胞群体。因此，iCT 实验允许对侵袭性细胞和非侵袭性细胞进行比较分析。
