Human autologous serum obtained using a completely closed bag system as a substitute for foetal calf serum in human mesenchymal stem cell cultures.
Mizuno N et. al. Cell Biology International (2006) Vol.30(3) pp.521-524

Increased proliferation of human synovial mesenchymal stem cells with autologous human serum.
Nimura A et. al. Arthritis & Rheumatism (2008) Vol.58(2) pp.501-510

Cytokine-rich autologous serum system for cartilaginous tissue engineering.
Isogai N et. al. Annals of Plastic Surgery (2008) Vol.60(6) pp.703-709

Serum preparation

Whole blood (70ml or 220ml) from 19 healthy donors was taken into CELLAID. After agitation, serum was separated by centrifuging the blood samples at 3,000×g for 7minutes. The volume of collected serum was measured, and the collection rate was calculated.

Measurement of coagulation markers and growth factors

The concentration of fibrinogen and the number of platelets in the blood samples which were being agitated were measured as coagulation markers. The concentrations of transforming growth factor beta1(TGF-beta1) , hepatocyte growth factor (HGF) and platelet-derived growth factor BB (PDGF-BB) were measured by ELISA method (R&D systems, USA).

In vitro culture of human mesenchymal stem cells (hMSCs)

Commercially available hMSCs (Lonza, USA) were seeded at the density of 500 cells/cm2 in MEM alpha (Gibco, USA) supplemented with 10 % of the serum prepared above. After 7 days , the numbers of cells was counted with coulter counter (Beckman coulter, USA).

40～60% of whole blood was obtained as serum regardless of collected blood volume.

Fibrinogen concentration and the number of platelets were decreased dramatically in first 20 minutes of agitation. (Figure 2.) The concentrations of growth factors reached plateau at 30 minutes of agitation.（Figure 3.)

“Growth Ratio” was calculated: the numbers of cells cultivated with each donor’s serum were divided by the average number of cells cultivated with 4 lots of FBS. Serum from the most of the donor had higher growth ratio than 1.0.