Please use this identifier to cite or link to this item: https://doi.org/10.1167/iovs.03-1361
Title: Development of a conjunctival epithelial equivalent with improved proliferative properties using a multistep serum-free culture system
Authors: Ang, L.P.K. 
Tan, D.T.H. 
Beuerman, R.W. 
Lavker, R.M.
Issue Date: Jun-2004
Citation: Ang, L.P.K., Tan, D.T.H., Beuerman, R.W., Lavker, R.M. (2004-06). Development of a conjunctival epithelial equivalent with improved proliferative properties using a multistep serum-free culture system. Investigative Ophthalmology and Visual Science 45 (6) : 1789-1795. ScholarBank@NUS Repository. https://doi.org/10.1167/iovs.03-1361
Abstract: PURPOSE. To investigate the use of a multistep serum-free culture system in developing a conjunctival epithelial equivalent with improved in vitro and in vivo proliferative properties and to evaluate the effect of serum supplementation and culture conditions on the proliferative capacity of these cells. METHODS. Conjunctival epithelial cells were cultivated on human amniotic membrane (HAM) in a multistep serum-free culture system, under submerged and air-lifted conditions. The bromodeoxyuridine (BrdU) ELISA proliferation assay, colony-forming efficiency (CFE), and number of cell generations were compared with those in serum-containing medium. The in vivo proliferative capability of the tissue-constructs were evaluated by xenotransplantation to SCID mice. Cultured cells were evaluated for the expression of keratin-4, -19, and -3, as well as MUC5AC goblet cell mucin. RESULTS. The epithelial cells cultivated in serum-free medium (BrdU absorbance, 1.91 ± 0.08; cell generations, 25.6 ± 4.5) were more proliferative than those cultivated in serum-containing medium (BrdU absorbance, 1.06 ± 0.08; cell generations, 12.1 ± 3.0). The serum-free-derived epithelial equivalents demonstrated a significant increase in proliferation and stratification after transplantation. Cells that were air lifted for 6 and 12 days had a reduced proliferative capacity in vitro and in vivo compared with submerged cultures. Cultured cells expressed keratin-4 and -19, and MUC5AC mRNA was detected by RT-PCR. Electron microscopy demonstrated a basal lamina with numerous hemidesmosomes. CONCLUSIONS. This is a multistep serum-free culture system for developing a conjunctival epithelial equivalent with improved proliferative and structural properties, which are crucial for enhancing graft survival and regeneration of the conjunctival surface after clinical transplantation.
Source Title: Investigative Ophthalmology and Visual Science
URI: http://scholarbank.nus.edu.sg/handle/10635/92635
ISSN: 01460404
DOI: 10.1167/iovs.03-1361
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.

SCOPUSTM   
Citations

49
checked on Sep 24, 2018

WEB OF SCIENCETM
Citations

45
checked on Sep 24, 2018

Page view(s)

47
checked on Sep 21, 2018

Google ScholarTM

Check

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.