Other cell types are fine - cardiac fibroblasts maintain the hydrogel for extended culture. When iPSC-CMs are seeded into the GelMA hydrogels, they are initially gelled (no movement of cells within, as seen under a microscope). After ~1hr the gels begin to disintegrate and are almost completely gone within 1 day.
It is clearly not inhibition of gelation, as the hydrogels are solid immediately after UV exposure. I don't know why iPSC-CMs would be so active as to degrade the gel within 1hr.
Dear Breanna Duffy san
Because it is simply natural gelatin, it is not safe. It is necessary to consider the test process from the raw material or the additive used or the substrate itself. In particular, we will promote the use of products that completely control the minerals and contamination of water used. Controls are what and how much is included.
Could it be because of any media supplement that you use for iPSC-CMs specifically? Have you checked this? Or if you do 3D cell culture so UV curing after cell seeding maybe UV exposure affects the membrane integrity in the short term have you checked the viability afterwards? Released products from cells could be degrading GelMA as it has a natural component. I would suggest checking the viability after an hour and do LDH assay to confirm damaged membrane integrity.
Thank you for the input! Yes, I checked the media - cell-free and cardiac fibroblast controls gel fine with the iPSC-CM media (RPMI + B27). Cell viability could definitely be a concern. There is a significant population of dead cells when harvesting the iPSC-CMs due to the shear stress of breaking the cells up (we can see it with trypan blue when doing a cell count). We have taken measures to reduce this as much as possible (ROCKi, DNAse during harvest), but still see significantly more dead cells than when harvesting other cell types that aren't as tightly connected. Any tips to further reduce cell death, or remove the dead cells before seeding in the hydrogel?
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