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<p>Secretome-based therapeutics offer promising cell-free approaches for regenerative medicine, yet current production methods face critical challenges in reproducibility, scalability, and product quality control. We developed an integrated microengineered platform combining electrospun polycaprolactone fibers with photopatterned polyethylene glycol diacrylate microwells for controlled adipose-derived stem cell spheroid formation and enhanced secretome production. The platform features 200&#xa0;μm microwells with 100&#xa0;μm spacing and an open-top, open-bottom architecture that enables bi-directional molecular diffusion. This design provides unrestricted nutrient transport to support spheroid viability while facilitating efficient secretome collection into culture medium. The platform generated uniform spheroids with diameters of 151.2 ± 11.3&#xa0;μm and minimal size variation (coefficient of variation: 7.5%), while maintaining cell viability exceeding 95% throughout extended culture periods. Comprehensive proteomic analysis using cytokine arrays revealed distinct secretory profiles between two-dimensional monolayer and three-dimensional spheroid cultures, with 37 proteins showing significant upregulation in spheroid-derived secretomes. Enzyme-linked immunosorbent assay quantification demonstrated a&#xa0;1.2-fold increase in epidermal growth factor and a&#xa0;1.8-fold increase in keratinocyte growth factor in spheroid cultures compared to monolayer controls. These results establish that identical cells cultured with identical media produce qualitatively different secretomes depending on culture platform architecture, demonstrating platform-dependent modulation of cellular secretory behavior. The integrated fabrication approach provides scalability through electrospinning technology and design flexibility through photolithographic patterning, addressing critical bioprocess engineering requirements for therapeutic secretome production. This platform offers a reproducible system for generating quality-enhanced secretomes with superior regenerative potential for tissue engineering and wound healing applications.</p>

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Electrospun fiber-based microwell platform for enhanced production of growth factor-enriched secretome from adipose stem cell spheroids

  • 期刊论文

  • 发表日期：

    2026年1月28日

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  • Dohyun Kim,
  • Hye Jin Hong,
  • Won-Gun Koh,
  • Hyun Jong Lee

摘要

Secretome-based therapeutics offer promising cell-free approaches for regenerative medicine, yet current production methods face critical challenges in reproducibility, scalability, and product quality control. We developed an integrated microengineered platform combining electrospun polycaprolactone fibers with photopatterned polyethylene glycol diacrylate microwells for controlled adipose-derived stem cell spheroid formation and enhanced secretome production. The platform features 200 μm microwells with 100 μm spacing and an open-top, open-bottom architecture that enables bi-directional molecular diffusion. This design provides unrestricted nutrient transport to support spheroid viability while facilitating efficient secretome collection into culture medium. The platform generated uniform spheroids with diameters of 151.2 ± 11.3 μm and minimal size variation (coefficient of variation: 7.5%), while maintaining cell viability exceeding 95% throughout extended culture periods. Comprehensive proteomic analysis using cytokine arrays revealed distinct secretory profiles between two-dimensional monolayer and three-dimensional spheroid cultures, with 37 proteins showing significant upregulation in spheroid-derived secretomes. Enzyme-linked immunosorbent assay quantification demonstrated a 1.2-fold increase in epidermal growth factor and a 1.8-fold increase in keratinocyte growth factor in spheroid cultures compared to monolayer controls. These results establish that identical cells cultured with identical media produce qualitatively different secretomes depending on culture platform architecture, demonstrating platform-dependent modulation of cellular secretory behavior. The integrated fabrication approach provides scalability through electrospinning technology and design flexibility through photolithographic patterning, addressing critical bioprocess engineering requirements for therapeutic secretome production. This platform offers a reproducible system for generating quality-enhanced secretomes with superior regenerative potential for tissue engineering and wound healing applications.