I culture animal chondrocytes (rabbit and porcine) and I would like to:
a) replace FBS in the culture media with allogenous serum
b) use serum-free medium with supplements.
Is there available any complete SF medium for chondrocytes to buy or do I have to mix it on my own?
If I have to mix it, I will supplement the basal medium (DMEM/F-12+GlutaMax) with ITS, dexamethasone, linoleic acid, ascorbic acid, growth factor and probably BSA. Unfortunatelly some of these components (like linoleic acid) are poorly or nonsoluble. Can you give me any advice or tips about mixing these components and about SF medium in general?
For FBS-free cultivation I will use cells, which were cultured in a culture medium supplemented with 10% FBS until second passage and frozen at - 79 °C.
all of the components which you want to add can be ordered already prepared for tissue culture. I use DMEM, high-glucose, with glutamate, HEPES, and non-essential amino acids. you can add FBS and other defined components as you wish.
when you revive your frozen cells, you should allow them to recover in 10-15 % FBS media, then use them for experiments (ie, plate them, remove FBS, etc).
I used Peprotech serum replacement with my primary mouse embryonic fibroblasts and it worked amazingly (we use FBS in are normal media). You could try that.
Hi, first I would like you to know that chondrocytes behave different in monolayer ordinary culture and no way to compare them with other cell type. It is known that chondrocyte losse their characterstics on monolayer culture. I developed a system of 3-D culture that keep the features of cartilage in vitro (Ahmed YA etal 2007). Second I found that serum type has little effect on the growth of chondrocytes. I used low FCS (0.1% + T3) to induce hypertropy and death as seen in vivo. low serum concentration alone (01%) keep the chondrocyte viable for longer time in culture but with no hypertophy. I hope I could help and I mention you to go through my papers for more detalis.
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