TY - JOUR
T1 - A virally inactivated functional growth factor preparation from human platelet concentrates
AU - Su, C. Y.
AU - Kuo, Y. P.
AU - Lin, Y. C.
AU - Huang, C. T.
AU - Tseng, Y. H.
AU - Burnouf, T.
PY - 2009/8
Y1 - 2009/8
N2 - Background Human platelet growth factors (HPGF) are essential for tissue regeneration and may replace fetal bovine serum (FBS) in cell therapy. No method for the manufacture of standardized virally inactivated HPGF has been developed yet. Study design and methods Platelet concentrates (PC) were subjected to solvent/detergent (S/D) treatment (1% TnBP/1% Triton X-45), oil extraction, hydrophobic interaction chromatography and sterile filtration. Platelet-derived growth factor (PDGF)-AB, -BB and -AA, transforming growth factor-ß1 (TGF-ß1), epidermal growth factor (EGF), insulin-like growth factor-1 (IGF-1) and vascular endothelium growth factor (VEGF) were measured by ELISA. Composition in proteins and lipids was determined, protein profiles were obtained by SDS-PAGE, and TnBP and Triton X-45 were assessed by gas chromatography and high-performance liquid chromatography, respectively. Cell growth promoting activity of HPGF was evaluated by 3-(4,5-dimethylthiazol-2-yl)- 5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay using human embryonic kidney (HEK293A) fibroblast and Statens Seruminstitute rabbit corneal (SIRC) epithelial cell lines. Results The GF preparation contained a mean of 16·66, 2·04, 1·53, 72·19, 0·33, 48·59 and 0·44 ng/ml of PDGF-AB, -BB, -AA, TGF-ß1, EGF, IGF-1 and VEGF, respectively. The protein profile was typical of platelet releasates and had less than 2 p.p.m. of residual S/D agents. MTS assay of HEK293A and SIRC cultures showed that the GF preparation at 10% and 0·1% (v/v), respectively, could successfully replace 10% FBS for cell proliferation. Cell-stimulating activity of HPGF on HEK293A was over twice that of PC releasates. Conclusion Standardized and functional virally inactivated HPGF can be prepared from human PC for possible applications in cell therapy and regenerative medicine.
AB - Background Human platelet growth factors (HPGF) are essential for tissue regeneration and may replace fetal bovine serum (FBS) in cell therapy. No method for the manufacture of standardized virally inactivated HPGF has been developed yet. Study design and methods Platelet concentrates (PC) were subjected to solvent/detergent (S/D) treatment (1% TnBP/1% Triton X-45), oil extraction, hydrophobic interaction chromatography and sterile filtration. Platelet-derived growth factor (PDGF)-AB, -BB and -AA, transforming growth factor-ß1 (TGF-ß1), epidermal growth factor (EGF), insulin-like growth factor-1 (IGF-1) and vascular endothelium growth factor (VEGF) were measured by ELISA. Composition in proteins and lipids was determined, protein profiles were obtained by SDS-PAGE, and TnBP and Triton X-45 were assessed by gas chromatography and high-performance liquid chromatography, respectively. Cell growth promoting activity of HPGF was evaluated by 3-(4,5-dimethylthiazol-2-yl)- 5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay using human embryonic kidney (HEK293A) fibroblast and Statens Seruminstitute rabbit corneal (SIRC) epithelial cell lines. Results The GF preparation contained a mean of 16·66, 2·04, 1·53, 72·19, 0·33, 48·59 and 0·44 ng/ml of PDGF-AB, -BB, -AA, TGF-ß1, EGF, IGF-1 and VEGF, respectively. The protein profile was typical of platelet releasates and had less than 2 p.p.m. of residual S/D agents. MTS assay of HEK293A and SIRC cultures showed that the GF preparation at 10% and 0·1% (v/v), respectively, could successfully replace 10% FBS for cell proliferation. Cell-stimulating activity of HPGF on HEK293A was over twice that of PC releasates. Conclusion Standardized and functional virally inactivated HPGF can be prepared from human PC for possible applications in cell therapy and regenerative medicine.
KW - Cell cultures
KW - Growth factors
KW - MTS assay
KW - Platelet
KW - Viral inactivation
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U2 - 10.1111/j.1423-0410.2009.01180.x
DO - 10.1111/j.1423-0410.2009.01180.x
M3 - Article
C2 - 19320900
AN - SCOPUS:68249133730
SN - 0042-9007
VL - 97
SP - 119
EP - 128
JO - Vox Sanguinis
JF - Vox Sanguinis
IS - 2
ER -