We have seeded these cell line within HA hydrogel and since the keratinocytes have formed spheroids overnight (50000 per well) we were thinking that it’s unlikely these spheroids are generated from singles cell. It seems that keratinocytes are able to migrate within the matrix to aggregate to each other. Therefore we are looking in literature for differences between spheroids generated from single cell and multi-cells.
May someone can share his/her experience or background?
Hi Adrien,
I work with fibroblasts and macrophages in 3D collagen gels. Our cells definitely can migrate within the matrix, but clumping seems cell type-specific, namely fibroblasts stay dispersed, but macrophages are more sociable and move towards other cells. I have never seen spheroids, though. If it is possible using your system, time lapse imaging of the cells would give you the clearest answer regarding how the spheroids form. Otherwise, maybe you could try to repeat the experiment using cells expressing different fluorescent markers.
Hi everybody,
As you mentionned and in order to see how keratinocytes migrate inside of the scaffold it couls be possible to perform fluorescence videomicroscopy. Moreover, in order to understand migration of keratinocyte one of the possibility will be to inhibite some migration or pathway or cell/cell interaction (e.g.cadherin interaction).
Hi Adrien,
We have worked much with keratinocytes inside collagen gel. First, it is very diffuclt to get single-cell suspension. So, cell clumps may be the source of spheroids. Second, keratinocytes are capable of morphogenesis in 3D cultures, including spheroids and tubules. See, for example http://link.springer.com/article/10.1007%2Fs00418-010-0691-0