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Human blood-derived fibrin releasates: Composition and use for the culture of cell lines and human primary cells
Authors:Thierry Burnouf  Chang-Yu Lee  Chin-Wan LuoYa-Po Kuo  Ming-Li ChouYu-Wen Wu  Yu-Hong TsengChen-Yao Su
Affiliation:a Human Protein Process Sciences (HPPS), Research Department, Lille, France & College of Oral Medicine, Taipei Medical University, Taipei, Taiwan
b School of Dentistry, College of Oral Medicine, Taipei Medical University & Division of Periodontics, Department of Dentistry, Taipei Medical University Hospital, Taipei, Taiwan
c Division of Oral and Maxillofacial Surgery, Department of Dentistry, Shuang Ho Hospital, Taipei Medical University, Taipei, Taiwan
d Gwo-Rei Biomedical Technology Group, Sanchong City, Taiwan
e Department of Microbiology & Immunology, Taipei Medical University, Taipei, Taiwan
f Gwo-Rei dental implant center, Biotechnology Department, Taipei, Taiwan
g Department of Dentistry, National Yang-Ming University, Taipei, Taiwan
Abstract:We have evaluated the capacity of two human blood fractions to substitute for FBS as growth medium supplement for human and animal cell cultures. Non-anticoagulated blood from volunteer donors (N = 13) was centrifuged to isolate a supernatant serum (SS) and a platelet-rich fibrin (PRF) clot which was squeezed to extract the releasate (PRFR). Both materials were characterized for the content in PDGF-AB, TGF-β1, VEGF, bFGF, EGF, IGF, total protein, albumin, IgG, IgM IgA, fibrinogen, cholesterol, triglycerides, various chemistry analytes and hemoglobin. Cell growth promoting activity of pooled SS and PRFR at 1, 5, and 10% in growth medium was evaluated over 7 days using human (HEK293, MG-63) and animal (SIRC, 3T3) cell lines and two human primary cultures (gingival fibroblasts and periodontal ligaments). Viable cell count was compared to that in cultures in FBS free-medium and 10% FBS supplement. SS and PRFR at 1-10% stimulated cell growth significantly more than FBS-free medium and in a way similar to 10% FBS in all cultures apart from 3T3. These two human blood-derived fibrin releasates are equally efficient to substitute for FBS as supplement for cell cultures and could be useful for specialized applications in regenerative medicine, dentistry and oral implantology, or cell therapy.
Keywords:Growth factors   Platelet   Blood   Cell culture   FBS
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