Zeitschrift für Bluterkrankungen und Transfusionen

Abstrakt

Serum Produced from Expired Fresh Frozen Plasma and Cryosupernatant Supports the Proliferation of Human Cells: Cost-Effective Alternatives to Fetal Bovine Serum

Kazuki Ishiyama, Mika Ogawa, Hidefumi Kato, Kyosuke Takeshita, Ryuzo Ueda, Takayuki Nakayama*

Background: Fetal Bovine Serum (FBS) is the most common growth factor used in cell culture. However, demands for xeno-free cell culture supplements for clinical use that comply with good manufacturing practice standards are increasing as cellular therapies expand. Human serum has been proposed as an alternative to FBS. Due to limited availability of donors and effective use of medical resources, we attempted to produce serum from expired Fresh Frozen Plasma (FFP) and Cryosupernatant (CS) and tested them as FBS substitutes.

Results: We found that 1-hour incubation of FFP or CS with a physiological level of thrombin and alcium Chloride (CaCl₂) depleted virtually all fibrinogen. Comparative proliferation studies with FBS and human Platelet Lysates (hPLs) showed that compared with FBS, FFP-serum exhibited similar or greater proliferative effects on 8 human cell lines: Bone Marrow-Derived Mesenchymal Stromal Cells (BMSC), Adipose Tissue-Derived Mesenchymal Stromal Cells (ADSC), HeLa, 293T, MG-63, HL-60, K562, and Meg-A2 except for Saos-2 cells, whereas CS-serum exhibited weaker proliferative effects on BMSC, ADSC and Saos-2 cells. hPL promoted the growth of BMSC and ADSC cells more vigorously than FFP- and CS-serum.

Conclusions: FFP-and CS-serum can be produced with thrombin and CaCl₂ rapidly and in a blood product–sparing manner and function as potent alternatives to FBS for culturing human cells.