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Preparation of Pooled Human Platelet Lysate pHPL as an Efficient Supplement for Animal Serum-Free Human Stem Cell Cultures
Published on: October 30, 2009
Plasma-Depleted Lyophilized Porcine Platelet Lysate as an Alternative to Fetal Bovine Serum in Cell Culture
Kuo-Chung Cheng1, Hung-Maan Lee1, Yi-Ting Shu2
1Department of Orthopedics, Taoyuan Armed Forces General Hospital, No. 168, Zhongxing Rd., Longtan Dist., Taoyuan City 325208, Taiwan.
Purpose:
Fetal bovine serum (FBS) is widely used in cell culture due to its rich nutrient and growth factor content, but it poses ethical concerns, biosafety risks, and cost limitations. This study investigates plasma-depleted lyophilized (freeze-dried) porcine platelet lysate (pPL) as a potential alternative to FBS for use in cell-based research and biomanufacturing.
Materials And Methods:
Fresh porcine blood was processed to obtain plasma-depleted pPL using double centrifugation and repeated freeze-thaw cycles. The lysate was analyzed for growth factor content via ELISA, then freeze-dried and sterilized with gamma irradiation. Endotoxin levels and cytotoxicity were evaluated. The ability of plasma-depleted lyophilized pPL to promote cell proliferation was assessed using L929 fibroblast cultures and compared with FBS.
Results:
Plasma-depleted lyophilized pPL contained significantly higher levels of TGF-β1 than FBS. The freeze-dried product remained stable for at least three months at room temperature. Gamma irradiation effectively sterilized the lysate without degrading key growth factors. Plasma-depleted lyophilized pPL showed no cytotoxicity and promoted greater proliferation of L929 cells compared to FBS, indicating enhanced mitogenic activity.
Conclusions:
Plasma-depleted lyophilized pPL is a stable, safe, and growth factor-rich alternative to FBS. It supports fibroblast proliferation, retains bioactivity after sterilization and storage, and may provide a scalable, ethical option for cell culture in biomedical research, regenerative medicine, and therapeutic product development.
