Making of Sheet of Culture Corneal Epithelium and Keratocyte by Thermal-response Cell Culture Dish.

2003 Fiscal Year Final Research Report Summary

• Project/Area Number: 14571688
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Ophthalmology
• Research Institution: Tokyo Women's Medical University, School of Medicine
• Principal Investigator: SHINOZAKI Kazurmi Tokyo Women's Medical University, School of Medicine, Department of Ophthalmology, Assistant, 医学部, 助手 (10256521)
• Co-Investigator(Kenkyū-buntansha): HORI Sadao Tokyo Women's Medical University, School of Medicine, Department of Ophthalmology, professor, 医学部, 教授 (20143498) ; TAKAMURA Etsuko Tokyo Women's Medical University, School of Medicine, Department of Ophthalmology, assistant professor, 医学部, 助教授 (90197202) ; YAMATO Masayuki Tokyo Women's Medical University, School of Medicine, Institute of Advanced Biomedical Engineering and Science, assistant professor, 医学部, 助教授 (40267117)
• Project Period (FY): 2002 – 2003
• Keywords: Thermal-resoonse Cell Culture Dish / Sheet of Culture Corneal Epithelium / Sheet of Culture Keratocyte

Research Abstract

The biomedical engineering research at our University has developed the thermal-response culture dish. The technique allows the collection of cell culture on a sheet without trypsins. With this method, we reproduced a corneal epithelium and a keratocyte layer with corneal epithelial cells and keratocyte cells. The sheets of corneal epithelial cells have so far been made with 3T3 cells as the feeder. However, in clinical applications, cultures from homogenous cells or serum are more preferable. We compared the reproduced corneal epithelium layers of white rabbits and of humans using : 1)keratocytes as the feeders as compared with the standard 3T3 cells ; 2)respective homogenous serum added to culture solutions as compared with the standard FBS ; and 3)different amount of growth factors. Similarly, reproduced keratocyte layers of white rabbits and humans using respective homogenous serum added to culture solutions as compared with the standard FBS were examined. The cell sheets made were examined under an optical microscope and a transmission electron microscope. The reproduced corneal epithelial cell sheets and the keratocyte sheets were transplanted to a white rabbit. The procedure allowed us to collect a reproduced keratocyte sheet similar to the collection of a 3T3 cell sheet. Although a cell layer could be reproduced successfully, a clear histological division of epithelium and other corneal layers as seen in a real cornea was not found. Serum reproduction was possible in both FBS and other homogenous serums. Whether a different amount of growth factor added to the culture solution could make a difference was unclear. At present, a reproduced epithelium layer and a keratocyte layer could be transplanted to the cornea of a white rabbit. However whether a reproduced keratocyte layer can substitute the original substance is unclear.