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Background <p>Animal cell culture has become an essential tool in biomedical research and industrial vaccine and biopharmaceutical development. Fetal bovine serum (FBS) is widely used as a culture supplement because of its rich composition of nutrients and growth factors. However, concerns over pathogen contamination and immunogenicity have prompted regulatory agencies to discourage its clinical application and motivated the development of serum-free media alternatives.</p> Results <p>In this study, a serum-free medium was developed for human lung fibroblasts (MRC-5) and optimized using response surface methodology in terms of the growth factor contents: EGF (9.3 ng/mL), IGF-1 (4.0 ng/mL), IGF-2 (4.3 ng/mL), ITS (4.5&#xa0;µg/mL), and PDGF-CC (6.9 ng/mL). The optimized growth factor mixture (OGM) (G2/M, 32.0%) and addition of 200&#xa0;µg/mL of <i>Haematococcus pluvialis</i> extracts (HE200) (OGM + HE200, G2/M, 35.3%) significantly enhanced cell proliferation by promoting cell cycle progression, particularly the G2/M phase compare with DMEM (G2/M, 15.0%). Moreover, Ki-67, a nuclear proliferation marker, showed increased expression in OGM-treated cells (mRNA, 2.7 fold) compared with those cultured in DMEM as evaluated by immunocytochemistry and qPCR. The addition of HE200 (mRNA, 4.0 fold) exerted synergistic effects with OGM, further promoting cell proliferation and the expression of cell cycle–related genes.</p> Conclusions <p>The study demonstrated the potential of combining an OGM and HE to support cell growth under serum-free conditions, providing a functional alternative to FBS.</p>

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Optimization of serum-free medium supplemented with microalgal extract using response surface methodology for human fibroblast culture

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年1月23日

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  • Eun-Jeong Koh,
  • Won-Kyu Lee,
  • Seong-Yeong Heo,
  • Areumi Park,
  • Eunyoung Jo,
  • Woon-Yong Choi,
  • Soo-Jin Heo

摘要

Background

Animal cell culture has become an essential tool in biomedical research and industrial vaccine and biopharmaceutical development. Fetal bovine serum (FBS) is widely used as a culture supplement because of its rich composition of nutrients and growth factors. However, concerns over pathogen contamination and immunogenicity have prompted regulatory agencies to discourage its clinical application and motivated the development of serum-free media alternatives.

Results

In this study, a serum-free medium was developed for human lung fibroblasts (MRC-5) and optimized using response surface methodology in terms of the growth factor contents: EGF (9.3 ng/mL), IGF-1 (4.0 ng/mL), IGF-2 (4.3 ng/mL), ITS (4.5 µg/mL), and PDGF-CC (6.9 ng/mL). The optimized growth factor mixture (OGM) (G2/M, 32.0%) and addition of 200 µg/mL of Haematococcus pluvialis extracts (HE200) (OGM + HE200, G2/M, 35.3%) significantly enhanced cell proliferation by promoting cell cycle progression, particularly the G2/M phase compare with DMEM (G2/M, 15.0%). Moreover, Ki-67, a nuclear proliferation marker, showed increased expression in OGM-treated cells (mRNA, 2.7 fold) compared with those cultured in DMEM as evaluated by immunocytochemistry and qPCR. The addition of HE200 (mRNA, 4.0 fold) exerted synergistic effects with OGM, further promoting cell proliferation and the expression of cell cycle–related genes.

Conclusions

The study demonstrated the potential of combining an OGM and HE to support cell growth under serum-free conditions, providing a functional alternative to FBS.