Hi there,

We used DMEM + 10% FBS and it worked just fine. However, during the first couple of hours we used a higher dose of FBS to facilitate cell attachment and after nearly 4 hours or so, the medium was replaced with a 10% FBS medium.

Thanks Hanie for the reply.

Could u please explain what is high dose FBS means ( ie 15 or 20 % FBS or more than this)???

Haven't worked with rabbit MSC but have worked extensively with human and mouse MSC. Check your serum. Some serum lots are better than others. These days you have MSC graded serums. Ask the vendor to get you a small sample to evaluate. Insist on checking the serum before ordering. Try isolating MSCs from younger animals. MSC seems to grow faster from young subjects. I always felt alpha MEM was way better than DMEM. So isolate cells from young animals, stick to alpha MEM and evaluate different serum at 15% dose. You can evaluate dose once you have got the optimum serum source.

Another way is to screen the literature, make an excel and see what are the common media component as given in different papers. I did this when I was confused for MSC differentiation cocktail. And then I read what the role of each of those components in differentiation and made a good differentiation cocktail.

Hope it helps.

Best.

50% FBS

Hello

DMEM/ low glucose supplemented with 10% fetal calf serum (FCS), 10 U/mL penicillin G, 10 μg/mL streptomycin, 2 mM L-glutamine .

also , i agree with Nishanth Gabriel for  alpha MEM

Good Luck

Thanks Saleh/Nishanth for reply

I started my first rabbit MSCs culture using DMEM (low glucose) after seeing literature suggested low glucose helps to maintain the stemness of the cells. Unfortunately it didnt worked for rabbit MSCs however the same media we used for human bone marrow MSC culture was doing so good...

As said earlier. Cells are getting adhered to the flask,but not proliferated for an week (increasing FBS concentration to 20% didnt helped). Later i added the DMEM/F12 then the cells reached confluent in a week. But the issue is in second passage itself there are senescence big cells.

In our experience FGF-2 supplementation is crucial. Rabbit MSC could be isolated reproducibly from bone marrow of 7 different NZW rabbits and grew very robustly simply with alfa-MEM medium, 10% FBS and 5 ng/ml FGF-2.

Also, regarding senescence, FGF-2 has a key role in delaying senescence and maintaining differentiation potential during expansion. More details on this point can be found in this couple of references (based on work with human cells, but I'm not aware of similar studies having been performed with rabbit cells):

http://www.ncbi.nlm.nih.gov/pubmed/12799186

http://www.ncbi.nlm.nih.gov/pubmed/22495904

Hope this helps,

Andrea

Dr.Banfi

Thanks for the reply...

I will give a try with 5 ng/ml  so far was using 1ng/ml FGF for rabbit MSCs...