Look good, let them grow a couple of days and you may have a full plate

Dear Min Ku Kang,

If I understood correctly, your fibroblast cells are looking “strange”; this might be usual, depending where these cells were isolated (tissue of origin). After isolation or cell passage, cells may display a different morphology and phenotype. If you’re using human cells, for instance, depending on age, cells may have different morphologies. Your culture seems to be at an early stage, with fibroblast at different stages of differentiation.

An important thing to consider is that usually cells may lose their ability to proliferate and to differentiate with cell passage, which is a well-known in vitro model for cell ageing; from my experience, aged cells look different, losing this “fibroblast-like” morphology. If you are using cells directly isolated from a tissue (explant, for example), there might be other non-adherent cells attached to your fibroblasts (blood cells?), that may give the impression that their morphology is looking odd. Usually, these might be easily removed with culture medium or with PBS (wash), unless they are connected through gap-junctions!...

Another point is the presence of these “bubbles”. These may, in fact, be apoptotic or not. Different works, including ours, have shown that membrane mechanism “blebbing”, which is reversible, is important during cell differentiation. For example, we have seen it in mesenchymal stem cells undergoing osteogenic differentiation, a mechanism that relies on Rho-kinase.

Hope it helps.

Regards.

Hi Min, I have to agree Casimiro. These fibroblasts look fine.