科研之友
微信
新浪微博
Facebook
分享链接
摘要
BACKGROUND: Human platelet concentrates (PCs) may be a source material to produce purified growth factors (GFs) for clinical use or cell therapy. However, no fractionation process of therapeutic-grade GF from PCs has ever been developed.
STUDY DESIGN AND METHODS: PCs were virally inactivated by solvent/detergent (S/D) treatment, subjected to oil extraction to remove part of the S/D agents, and fractionated on a SP-Sepharose (SP) matographic column equilibrated in a phosphate-buffered saline (PBS) buffer, pH 7.5. The breakthrough was recovered, and the column was washed with the PBS buffer and then eluted by a 0.7 mol/L NaCI-PBS buffer pH 7.5 (SP-eluate). The SP-breakthrough and SP-eluate were characterized for their content in GF, proteins, lipids, and S/D agents. The MTS value of three cell lines cultivated in a medium containing 10% fetal bovine serum supplemented with 1% to 3% of SP-eluate or recombinant human (rHu) platelet-derived growth factor (PDGF)-BB was compared.
RESULTS: The SP-eluate contained a mean of 47, 17, and 6 ng/mL PDGF-AB, -BB, and -AA, respectively, 0.26 ng/mL vascular endothelial growth factor (VEGF). It was largely depleted of transforming growth factor-beta 1 (2.33 ng/mL), epidermal growth factor (0.09 ng/mL), insulin-like growth factor (3.40 ng/mL), albumin, immunoglobulin (Ig)G, IgM, IgA, and fibrinogen, which were mostly in the breakthrough. tri-n-butyl phosphate and Triton X-45 were less than 2 ppm. Cell growth promoting activity of the SP-eluate was at least as as that of rHu-PDGF-BB.
CONCLUSION: Human PC can be fractionated into a purified, virally inactivated PDGF and VEGF concentrate, opening perspectives for the development of a new range of blood products for clinical use and cell therapy procedures.
- 出版日期2010-8
