N-cadherin-mediated aggregate formation; cell detachment by Trypsin-EDTA loses N-cadherin and delays aggregate formation

Abstract Although cell aggregates/spheroids are useful tools in various fields of cell biology, the mechanism for aggregate formation is not fully resolved yet. Here I show the involvement of N-cadherin in the quick formation of packed aggregates in suspension culture. HEK293T cells detached from substratum by Trypsin alone quickly formed packed aggregates in suspension. This aggregate formation was inhibited by the down-regulation of N-cadherin. Meanwhile, aggregate formation of cells detached by Trypsin-EDTA was significantly delayed. N-cadherin was transiently lost by Trypsin-EDTA-treatment, and the re-expression of N-cadherin corresponded to delayed aggregate formation. Furthermore, packed phenotype was not observed in the absence of N-cadherin. These findings indicate that N-cadherin mediates quick formation of packed aggregates/spheroids in suspension culture.