Structural characterization of bioengineered human corneal endothelial cell sheets fabricated on temperature-responsive culture dishes
2005; Elsevier BV; Volume: 27; Issue: 4 Linguagem: Inglês
10.1016/j.biomaterials.2005.06.005
ISSN1878-5905
AutoresTakeshi Ide, Kohji Nishida, Masayuki Yamato, Taizo Sumide, M Utsumi, Takayuki Nozaki, Akihiko Kikuchi, Teruo Okano, Yasuo Tano,
Tópico(s)Ocular Surface and Contact Lens
ResumoFor the purpose of corneal regenerative medicine, we fabricated human corneal endothelial cell sheets on temperature-responsive dishes, which could be non-invasively harvested as intact, transplantable sheets by simply reducing the culture temperature. Cells demonstrated hexagonal cell shape with numerous microvilli and cilia, and also exhibited abundant cytoplasmic organelles similar to these cells in vivo. Immunofluorescence for type IV collagen and fibronectin revealed that abundant extracellular matrix (ECM) was deposited on the basal surface throughout culture, and the deposited ECM was harvested along with the cell sheets by reducing culture temperature to 20 °C. Faint ECM remnants were observed on the dish surfaces after cell sheet detachment. Immunofluorescence for ZO-1 showed that tight junctions were established between cells, and immunoblotting indicated that intact ZO-1 was maintained during cell sheet harvest, while conventional proteolytic cell harvest methods resulted in the degradation of ZO-1. These results suggest that these transplantable corneal endothelial cell sheets can be applied to treat patients with damaged corneas.
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