I'm starting some work with human mesenchymal stem cells and wondering what media can be used for general culture in their non-differentiated state? I'm purchasing the cells from ATCC and can buy the growth media to go with the cells, however it works out to be very expensive buying their basal media with the additional growth kit (~£170 per bottle of media). Just checking if there are any as-good cheaper alternatives?
In my experience with mesenchymal stem cells from umbilical cord tissue (Wharton jelly) they grow very well with DMEM low glucose with 10% serum or 10% platelet lysate. I’ve tested other mediums such as DMEM F12 or DMEM high glucose but they didn’t grow many passages. I don´t think you need any special media for growing these cells, DMEM low glucose will do just fine.
Diego Raffo - thank you for your response. I'll give low glucose DMEM, 10 % serum a try.
In my expirament whatron jelly explant extraction
We used
DMEM low glucose
with 20% FCS and 1% antibiotic
0.1% L.glutamine
DMEM with 10% FBS supplementation maintain a good proliferation of MSCs. You can supplement your cell culture medium with fibroblast growth factor 1-5 ng/ml (FGF) to enhance the proliferation without drastic phenotypic changes.
The classical MSC media has always been Alpha MEM with FBS. If using FBS batch test as many lots as you can, as the performance varies hugely between lots. aMEM with 10% FBS at minimum is a good starting point. Human platelet lysate is a good growth supplement if you can get it. Use at about 5-10% and add heparin or it will clot. The gold standard media are serum free or chemically defined as these have no lot to lot variability and tend to protect the 'stemness' of the cells over time. Stem Cell Technologies have a great CD media (Disclosure- I made it).
Do you know what is the source for those mesencymal cells? DMEM low glucose is the most frequent for umbilical cord tissue derived cells, however, from bone marrow alfa-MEM is very popular and IMDM or alpha-MEM for umbilical cord blood derived mesenchymal cells.
Dear Kathryn,
Our lab compared various media. For our MSCs, DMEM low glucose or alpha MEM + 10% in house processed platelet lysate + heparin to prevent clotting worked well. FBS supplementation caused early senescence, and chemically defined medium needs attachment factors, so needs an additional cost.
Thank you everyone for your helpful responses. Diego Raffo they are bone marrow derived MSCs. I'll try DMEM low glucose, 10 % FBS, 1 % antibiotic. If they're slow to start off I'll add glutamine.
If posible, life technologies sells medium with glutamax, and there are other similar alternatives of glutamin that don't degrade. They are simpler to use
I always use alpha MEM + 10% FBS +1% antibiotic for bone marrow-derived MSC. The grow up very well and quickly.
Hello
1. Dulbecco’s modified Eagle’s medium low glucose (DMEM-LG, Invitrogen, Karlsruhe, Germany) with 10% fetal calf serum (FCS, Biochrom, Berlin, Germany) and 1% penicilline/streptomycine (Invitrogen, Karlsruhe, Germany).
2. Alpha minimum essential medium (αMEM) with L-glutamine with 10% fetal calf serum (FCS, Biochrom, Berlin, Germany) and 1% penicilline/streptomycine (Invitrogen, Karlsruhe, Germany).
3. A variation of “Verfaillie” medium, consisting of 547.5 ml Dulbecco’s modified Eagle’s medium high glucose (DMEM-HG, Invitrogen, Karlsruhe, Germany) and L-glutamine (Invitrogen, Karlsruhe, Germany), 40 ml MCDB (Sigma-Aldrich, Steinheim, Germany), 20 ml FCS (Biochrom, Berlin, Germany), 10 ml penicilline/streptomycine (Invitrogen, Karlsruhe, Germany), 20 ml IST (Sigma-Aldrich, Steinheim, Germany), 2 ml dexamethasone (Sigma-Aldrich, Steinheim, Germany), 500 μl ascorbic acid (Sigma-Aldrich, Steinheim, Germany), 10 ng/ml PDGF-BB (Miltenyi Biotec, Bergisch Gladbach, Germany), 10 ng/ml EGF (Miltenyi Biotec, Bergisch Gladbach, Germany).
4. A variation of “Bernese chondrocyte medium”consisting of Dulbecco’s modified Eagle’s medium F12 + L-glutamine (Invitrogen, Karlsruhe, Germany), 1% penicilline/streptomycine (Invitrogen, Karlsruhe, Germany), 10% FCS (Biochrom, Berlin, Germany), 10 μl/50 ml TGF-ß1 (Acris, Herford, Germany), 2.5 μl/50 ml FGF-2 (Acris, Herford, Germany).
References
Article Different culture media affect growth characteristics, surfa...
Article Ex Vivo Expansion of Human Mesenchymal Stem Cells in Defined...
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