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Properties of Corneas Reconstructed with Cultured Human Corneal Endothelial Cells and Human Corneal Stroma

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Abstract

Purpose

To examine the properties of corneas tissue-engineered with cultured human corneal endothelial cells (HCEC) and human corneal stroma.

Methods

Primary HCEC cultures were established from endothelial cell layer explants and propagated on culture dishes coated with bovine corneal endothelial extracellular matrix. A cell suspension of HCEC at the fifth passage was transferred onto human corneal stroma deprived of endothelial cells, and the corneas were gently centrifuged to enhance cell attachment. The cell density of the tissue-engineered corneas was examined after staining with alizarin red and trypan blue. The tissue-engineered corneas were histologically examined by light and electron microscopy. The pump function of the tissue-engineered corneas was measured using an Ussing chamber.

Results

The mean endothelial cell density of four tissue-engineered corneas was 2380 ± 264 cells/mm2 (mean ± SD). HCEC on the tissue-engineered corneas had a morphology similar to HCEC in vivo. The pump function parameters of the tissue-engineered corneas were 55%–75% of those of normal corneas.

Conclusions

HCEC on the tissue-engineered corneas have morphology and cellular density similar to HCEC in vivo, whereas the pump function of the tissue-engineered corneas was lower than in normal corneas. Jpn J Ophthalmol 2005;49:448–452 © Japanese Ophthalmological Society 2005

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Authors and Affiliations

  1. Department of Ophthalmology, University of Tokyo School of Medicine, Tokyo, Japan

    Shiro Amano, Tatsuya Mimura & Satoru Yamagami

  2. Meiwakai Medical Foundation, Miyata Eye Hospital, Miyakonojo, Miyazaki, Japan

    Yasuhiro Osakabe & Kazunori Miyata

Authors
  1. Shiro Amano
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  2. Tatsuya Mimura
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  3. Satoru Yamagami
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  4. Yasuhiro Osakabe
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  5. Kazunori Miyata
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Corresponding author

Correspondence to Shiro Amano.

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Amano, S., Mimura, T., Yamagami, S. et al. Properties of Corneas Reconstructed with Cultured Human Corneal Endothelial Cells and Human Corneal Stroma. Jpn J Ophthalmol 49, 448–452 (2005). https://doi.org/10.1007/s10384-005-0245-5

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  • DOI: https://doi.org/10.1007/s10384-005-0245-5

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