A virally inactivated platelet-derived growth factor/vascular endothelial growth factor concentrate fractionated from human platelets.
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.


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) chromatographic 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 NaCl-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, and 0.26 ng/mL vascular endothelial growth factor (VEGF). It was largely depleted of transforming growth factor-β1 (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 good as that of rHu-PDGF-BB.


CONCLUSIONS
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.
Burnouf
Thierry
T
Human Protein Process Sciences, Research Department, Lille, France.
Kuo
Ya Po
YP
Huang
Chun Ting
CT
Tseng
Yu Hung
YH
Lin
Che Tong
CT
Su
Chen Yao
CY
D001792:Blood Platelets / Q000737:chemistry*; D002478:Cells, Cultured; D002845:Chromatography; D006801:Humans; D010976:Platelet Count; D010982:Platelet-Derived Growth Factor / Q000302:isolation & purification*; D042461:Vascular Endothelial Growth Factor A / Q000302:isolation & purification*
D010982:Platelet-Derived Growth Factor; D042461:Vascular Endothelial Growth Factor A