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

Thierry Burnouf, Ya Po Kuo, Chun Ting Huang, Yu Hung Tseng, Che Tong Lin, Chen Yao Su

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

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) 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. 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.

Original languageEnglish
Pages (from-to)1702-1711
Number of pages10
JournalTransfusion
Volume50
Issue number8
DOIs
Publication statusPublished - 2010

Fingerprint

Platelet-Derived Growth Factor
Sepharose
Vascular Endothelial Growth Factor A
Blood Platelets
Detergents
Intercellular Signaling Peptides and Proteins
Buffers
Phosphates
Cell- and Tissue-Based Therapy
Octoxynol
Transforming Growth Factors
Somatomedins
Epidermal Growth Factor
Fibrinogen
Immunoglobulin A
Immunoglobulin M
Albumins
Oils
Immunoglobulin G
Lipids

ASJC Scopus subject areas

  • Hematology
  • Immunology
  • Immunology and Allergy
  • Medicine(all)

Cite this

A virally inactivated platelet-derived growth factor/vascular endothelial growth factor concentrate fractionated from human platelets. / Burnouf, Thierry; Kuo, Ya Po; Huang, Chun Ting; Tseng, Yu Hung; Lin, Che Tong; Su, Chen Yao.

In: Transfusion, Vol. 50, No. 8, 2010, p. 1702-1711.

Research output: Contribution to journalArticle

Burnouf, Thierry ; Kuo, Ya Po ; Huang, Chun Ting ; Tseng, Yu Hung ; Lin, Che Tong ; Su, Chen Yao. / A virally inactivated platelet-derived growth factor/vascular endothelial growth factor concentrate fractionated from human platelets. In: Transfusion. 2010 ; Vol. 50, No. 8. pp. 1702-1711.
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abstract = "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) 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. 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.",
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T1 - A virally inactivated platelet-derived growth factor/vascular endothelial growth factor concentrate fractionated from human platelets

AU - Burnouf, Thierry

AU - Kuo, Ya Po

AU - Huang, Chun Ting

AU - Tseng, Yu Hung

AU - Lin, Che Tong

AU - Su, Chen Yao

PY - 2010

Y1 - 2010

N2 - 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) 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. 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.

AB - 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) 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. 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.

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