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Journal of Artificial Organs

, Volume 4, Issue 4, pp 353–356 | Cite as

Tissue-engineered skin using aggregates of normal human skin fibroblasts and biodegradable material

  • Katsuko S. Furukawa
  • Takashi Ushida
  • Yasuyuki Sakai
  • Kaiko Kunii
  • Motoyuki Suzuki
  • Junzo Tanaka
  • Tetsuya Tateishi
Original Article Artificial Skin

Abstract

Higher-density inoculation of fibroblasts into a three-dimensional scaffold should accelerate wound healing after skin implantation. This study attempted to develop tissue-engineered skin with a higher density of fibroblasts. We first attempted to fabricate three-dimensional high-cell-density aggregates (spheroids) of normal human fibroblasts for application to tissue-engineered skin. Our method consisted of rotational shaking with nontreated dishes, decreasing fibroblast-meterial interactions, and augmenting cell-cell interaction. To prompt aggregate formation, the medium was supplemented with insulin, dexamethasone, ascorbic acid, and basic fibroblast growth factors that potentiate secretion of extracellular matrices. Under such improved conditions, fibroblasts were able to form spheroidal aggregates within 24 to 36h of rotational culture. Although the formed aggregates were irregular in shape and were composed of only several cells after 12h, they became almost spheroidal after 24h. The aggregates grew even more round after 36h, and their surfaces became smooth. After 36h of rotational culture, the fibroblast aggregates were collected and reinoculated onto a biodegradable mesh composed of polyglycolic acid coated with collagen. The aggregates were trapped in the material and became attached after 24h. Finally, because transforming growth factor-β3 (TGF-β3) is known to accelerate wound healing, we conducted a semiquantitative analysis of TGF-β3 mRNA in both the fibroblast monolayers (two-dimensional culture) and the aggregates (three-dimensional culture). Analysis of TGF-β3 mRNA expression showed that mRNA expression was greater in the fibroblasts of aggregates than in a monolayer. Therefore, our newly developed dermal graft is expected to accelerate wound healing faster than conventional grafts.

Key words

Normal human skin fibroblast Biodegradable polymer Tissue-engineered skin 

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Copyright information

© The Japanese Society for Artificial Organs 2001

Authors and Affiliations

  • Katsuko S. Furukawa
    • 1
  • Takashi Ushida
    • 1
    • 2
  • Yasuyuki Sakai
    • 3
  • Kaiko Kunii
    • 2
  • Motoyuki Suzuki
    • 3
  • Junzo Tanaka
    • 4
  • Tetsuya Tateishi
    • 1
    • 2
  1. 1.Biomedical Engineering Laboratory (Tateishi & Ushida Laboratory), Department of Mechanical Engineering, Graduate School of EngineeringUniversity of TokyoTokyoJapan
  2. 2.Dimensional Cell & Tissue Engineering Group, National Institute for Interdisciplinary Research, Agency of Industrial Science and TechnologyMinistry of International Trade and IndustryTsukubaJapan
  3. 3.Institute of Industrial ScienceUniversity of TokyoTokyoJapan
  4. 4.National Institute for Research in Inorganic MaterialsTsukubaJapan

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