A virally inactivated functional growth factor preparation from human platelet concentrates

C. Y. Su, Y. P. Kuo, Y. C. Lin, C. T. Huang, Y. H. Tseng, T. Burnouf

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)119-128
Number of pages10
JournalVox Sanguinis
Volume97
Issue number2
DOIs
Publication statusPublished - Aug 2009

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Intercellular Signaling Peptides and Proteins
Blood Platelets
Vascular Endothelium
Octoxynol
Transforming Growth Factors
Somatomedins
Cell- and Tissue-Based Therapy
Epidermal Growth Factor
Detergents
Rabbits
Proteins
Regenerative Medicine
Serum
Hydrophobic and Hydrophilic Interactions
Human Activities
Gas Chromatography
Chromatography
Regeneration
Polyacrylamide Gel Electrophoresis
Oils

Keywords

  • Cell cultures
  • Growth factors
  • MTS assay
  • Platelet
  • Viral inactivation

ASJC Scopus subject areas

  • Hematology

Cite this

A virally inactivated functional growth factor preparation from human platelet concentrates. / Su, C. Y.; Kuo, Y. P.; Lin, Y. C.; Huang, C. T.; Tseng, Y. H.; Burnouf, T.

In: Vox Sanguinis, Vol. 97, No. 2, 08.2009, p. 119-128.

Research output: Contribution to journalArticle

Su, C. Y. ; Kuo, Y. P. ; Lin, Y. C. ; Huang, C. T. ; Tseng, Y. H. ; Burnouf, T. / A virally inactivated functional growth factor preparation from human platelet concentrates. In: Vox Sanguinis. 2009 ; Vol. 97, No. 2. pp. 119-128.
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abstract = "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-{\ss}1 (TGF-{\ss}1), 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-{\ss}1, 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.",
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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

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KW - Viral inactivation

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