I use 2% chitosan (w/v) along with 5.6% beta glycerophosphate (w/v) along with 0.1 M sodium bicarbonate. Cell concentration being 2x106 cells/ml. Previously i used MG63 cell lines and they used to show excellent proliferation but now these cells are simply migrating out of the scaffold.
Anything in the media to induce chemotaxis? Also is the scaffold becoming glucose depleted ? This could cause your observations…
William Richard Webb I change the media in every 48 hours and the media is normal stem cell culture media containing -
1. 5ml FBS
2. 45ml knock DMEM
3. 500ul Glutamax
4. 250ul antibiotic
5. 10ng/ml FGF
William Richard Webb No, fgf is only present in the stem cell culture media but media enters the scaffold because they turn pinkish red eventually?
Yes fgf in media can cause chemotaxis along with glucose concentration gradient between media and scaffold. I never used fgf for standard culture of MSCs unless they stopper proliferating
William Richard Webb Okay, I will try this once and get back to you. Thank you so much, for your input. But what can I do about glucose concentration gradient?
Nothing unless you use media in the fabrication of the hydrogel and the hydrogel allows diffusion
Got it.....Thanks a lot, Sir!!! I will give these a try and get back to you.
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