Dear Urban, basically the published papers indicate that you can use the same basal medium supplemented using platelet lysate as you would do using fetal bovine serum. In our case, we have experience in the use of DMEM-LG supplemented with 10% human platelet lysate for expanding human adipose-derived stem cells, and it worked quite well for expansion, morphology, doubling time, maintenance of phenotype over > 7 passages, and differentiation into the 3 typical cell lineages tested (chondrocytes, osteo, and adipocytes).  Best, Thierry  

i've used alpha-MEM for primary culture of UC-MSC and  DMEM-low glucose for further passaging. Also i've used PRP as serum substitute. 

Hello 

yes , if you need further passaging use alpha-MEM for primary culture and DMEM-low glucose .

Preparation of Supplements and Media :

- Dulbecco’s modified Eagle’s medium (DMEM) supplemented with either 10% MSC-qualified FBS or with 5% PLTMax supplement , 2 mM l-glutamine , 1× EmbryoMax penicillin– streptomycin solution , 8 ng/mL human recombinant basic fibroblast growth factor (bFGF) , and 2 U/mL heparin . Collagen-coated microcarriers were chosen for hMCs expansion in spinner flasks .

- Dulbecco’s modified Eagle’s medium-low glucose (DMEM-LG)

with 4 mM L-glutamine and 0.05 units of penicillin per 0.05 g of streptomycin (PAA)

served as the basal medium, which was supplemented with (a) 10%

fetal calf serum (preselected lot; mesenchymal stem cell growth

supplement; Cambrex), (b) 10% AB-HS, or (c) 10% tPRP.

In our lab we used alpha MEM supplemented by 10% PRP for BMP, adipose tissue and UC tissue derived MSC with good result

When you use platelet lysate, what is the number of platelets you use per militer of plasma before lysis?

The PRP is actually thrombocyte concentrate from Indonesian red cross. We have checked the platelet count, it is between 200,000 and 1,200,000. upon freeze thaw, not all the platelets were lyzed, as there was a residual number of platelets. We used one two or three freeze thaw cycles, and the growth factor content did not differ significantly between one, two or three cycles. We supposed that every cycle, a new batch was lyzed.  Therefore, when the thrombocyte count is high, more freeze thaw cycles can be done. Over expiry date PRP still contain growth factors and can be used as culture medium supplement. (Sari P, Luviah E, Nugraha Y, Pawitan JA, Lilianty J, Purwoko RY. Various growth factor yields in various platelet rich plasma processing methods. BTAIJ 2014; 9(11):475-478. Yudhi Nugraha, Puji Sari, Reza Yuridian Purwoko, Evah Luviah, Jeanne Adiwinata Pawitan. Effect of Lysed Platelet Count in Platelet Concentrates on Various Growth Factor Levels after Freeze Thaw Cycles. Int J PharmTech Res 2014; 6(7): 2036-42.)

We usually aliquot  the PRP in  5 mL tubes, and freeze, and used 1-3 mL each time. So, the freeze thaw cycles can be 1-3 times.