Interactions of B16F10 melanoma cells aggregated on a cellulose substrate
Interactions of B16F10 melanoma cells aggregated on a cellulose substrate
复制标题
DOI：
10.1002/jcb.20833
复制
发表时间：
2006-09-01
影响因子：
4
通讯作者：
Nagel, M. -D.
中科院分区：
文献类型：
--
作者：
Hindie, M.;Vayssade, M.;Nagel, M. -D.
有证据表明，细胞的形状及其与基质的接触直接影响正常细胞和癌细胞的生长和分化。 3D 培养中的细胞比传统 2D 培养中的细胞更接近体内情况。我们研究了 B16F10 小鼠黑色素瘤细胞的相互作用和功能，当这些细胞在纤维素包被的培养皿 (CEL) 上聚集时，它们在组织培养聚苯乙烯 (tPS) 上扩散和生长良好。 CEL 上黑色素瘤细胞的这种聚集是 Ca2+ 依赖性的，并由 N-钙粘蛋白介导。在 CEL 和 tPS 上培养的细胞中，N-钙粘蛋白和 P-连环蛋白转录物水平相似，但 CEL 上的细胞含有较少的 P-连环蛋白。免疫沉淀和免疫染色显示，CEL 上的细胞膜上存在 N-钙粘蛋白和 β-连环蛋白。在 CEL 上 48 小时后，细胞增殖明显减慢，并且纤维素涂层导致大多数细胞停滞在 G(1) 中。我们还比较了 CEL 上细胞和 tPS 上生长的对照细胞的黑色素含量和酪氨酸酶活性。在 CEL 上聚集的细胞中诱导黑色素生成。因此，纤维素基质似乎是研究 3D 培养物中细胞间相互作用和细胞功能的出色工具。
There is evidence that the shape of cells and their contact with a matrix direct the growth and the differentiation of both normal and cancer cells. Cells in 3D culture resemble the in vivo situation more closely than do those in conventional 2D cultures. We have studied the interactions and functions of B16F10 mouse melanoma cells, which spread and grow well on tissue culture polystyrene (tPS), when they were made to aggregate on cellulose-coated Petri dishes (CEL). This aggregation of melanoma cells on CEL was Ca2+ dependent and mediated by N-cadherins. The levels of N-cadherin and P-catenin transcripts in cells cultured on CEL and tPS were similar, but those on CEL contained less P-catenin protein. Immunoprecipitation and immunostaining showed that both N-cadherins and beta-catenins were present at the membranes of cells on CEL. Cells proliferated significantly more slowly after 48 h on CEL and the cellulose coating caused most of them to arrest in G(1). We also compared the melanin contents and tyrosinase activity of cells on CEL and controls grown on tPS. Melanogenesis was induced in cells aggregated on CEL. A cellulose substrate thus appears to be an outstanding tool for studying cell-cell interactions and cell functions in 3D cultures.