Hello all,
We cultured MSCs on calcium phosphate discs for 3 days and 7 days. We are seeing strange crystal-like precipitates or something of the sort (images attached). They are found wherever cells are found, or nearby cells, that are growing on the surface of the discs. We did EDS on these samples out of curiosity and the crystals appear to have a high concentration of NaCl, which indicates that they are salts.
I can't find any literature that shows this happening in their cell studies. Has anyone else seen this sort of thing happen in their cell cultures? I have no idea what could explain these results and I would appreciate some insights, or hypotheses, if any.
Thanks!
Hi Genesis Marrero
Interesting observation! Based on your description, it seems like the crystals could indeed be salt precipitates. There could be several reasons for this, here are a few possible explanations. Media evaporation: If your cultures are not fully sealed or the incubator is not properly humidified, evaporation could cause the salts in the culture medium to become more concentrated over time. This might lead to precipitation, especially near cells which could act as nucleation points for crystal formation. Interaction with the disc material: Calcium phosphate could be reacting with components of the culture medium, leading to formation of insoluble salts. I hope these ideas help you in understanding and investigating this phenomenon further!
A brief water soak before processing for critical point drying should get rid of the salts.
Not clear for me. You see these crystals already in your cell culture or only after SEM preparation?
Do you fix your samples with PBS for SEM preparation? PBS is very salty, you need very good washing before dehydration with solvent. Use sodium cacodylate to fix your sample.
Nicolas Barois Yes, we fix our samples with PBS using Karnovsky's fixative solution followed by a graded ethanol dehydration. We might try using sodium cacodylate but we were hoping to not have to use this.
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