How many cells should I seed in each well?
每个孔中应播种多少个细胞？
Aim for 95-100% confluent monolayers around the cell exclusion zone at the start of the experiment. Seeding at a higher density may result in cells in the Detection Zone at the beginning of the assay while seeding at a lower density will yield fewer migrating cells in the detection zone after migration. Typically, 20,000 – 75,000 cells are seeded per well in Oris and Oris Pro 96-well plates, and 2,500 – 10,000 per well in Oris Pro 384-well plates. For your first experiment, we recommend testing several different cell seeding densities as described in Appendix 1 in the assay protocol. For Oris 3D Embedded Invasion assays, we recommend seeding at 30,000 – 50,000 cells per well. As many as 75,000 cells per well may be seeded for some cell lines, but many invasive cells lines express high levels of matrix metalloproteinases that can degrade the collagen to the point where it collapses or even liquefies at high cell densities. Adding fewer cells ameliorates this problem.
实验开始时，细胞排斥区周围要有 95-100% 融合单层细胞。如果播种密度较高，可能会导致细胞在实验开始时出现在检测区，而如果播种密度较低，则细胞迁移后在检测区出现的迁移细胞较少。通常情况下，Oris 和 Oris Pro 96 孔板每孔播种 20,000 - 75,000 个细胞，Oris Pro 384 孔板每孔播种 2,500 - 10,000 个细胞。对于您的首次实验，我们建议您测试几种不同的细胞播种密度，如《Oris Pro》中附录 1 所述。 检测规程. 对于 Oris 3D 嵌入式侵袭检测，我们建议每孔播种 30,000 - 50,000 个细胞。某些细胞系每孔可播种多达 75,000 个细胞，但许多侵袭性细胞系表达高水平的基质金属蛋白酶，可降解胶原蛋白，使其在高细胞密度下塌陷甚至液化。减少细胞数量可改善这一问题。