Hello,
After seeding a homogeneously distributed cell culture on top of the surface of a (polyisocyanopeptide-) hydrogel, I find clustering of cells in many of the wells. In some (even adjacent ones) I see a confluent cell layer.
Does anyone have an idea what could be the reason for clustering? And what I could do to prevent it the next time?
I use the following:
- Vaginal fibroblasts, Passage 5 (have passaged it one time before in culture flask)
- 96 well plate (seeded appr 4000 cells p well)
- Polyisocyanopeptide diluted in medium as substrate (forms a soft gel). 80 ul p well
- Medium: DMEM/F-12 (88%), FBS (10%), pen/strep (2%)
I you need more info please let me know!
Thanks a lot for the help
Hi Aksel
have you ever figured out why did this type of cluster happen ? I have similar problem with fibroblasts when they are seeded on top of hydrogel !
Hi Karrar,
Thanks for your reply.
I was not yet familiar with cell culture using hydrogels when posting this question.
It is common for cells to grow in clusters when seeded on hydrogels.
My hypothesis is that it depends on the number and distribution of attachment sites available for the cells. If you are able to increase those, more clusters will appear, however spread over the surface of the substrate, interconnecting with one another. This increases cell growth.
What hydrogel are you using?
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