+ Site Statistics
References:
52,654,530
Abstracts:
29,560,856
PMIDs:
28,072,755
+ Search Articles
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ PDF Full Text
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Translate
+ Recently Requested

A novel tissue engineering approach for ocular surface reconstruction using bioengineered corneal epithelial cell sheet grafts from limbal stem cells expanded ex vivo on a temperature responsive cell culture surface



A novel tissue engineering approach for ocular surface reconstruction using bioengineered corneal epithelial cell sheet grafts from limbal stem cells expanded ex vivo on a temperature responsive cell culture surface



ARVO Annual Meeting Abstract Search & Program Planner : Abstract No 3149



Purpose: Previously we reported a cell sheet manipulation technique that utilizes temperature-responsive cell culture surfaces. Confluent cell cultures on these surfaces are harvested as a single contiguous cell sheet retaining cell-cell junctions as well as a deposited extracellular matrix on the basal sheet surface. In this study, we have developed a novel tissue engineering approach for ocular surface reconstruction using limbal stem cells expanded ex vivo on a temperature-responsive cell culture surface. Methods: Rabbit corneal limbal epithelial cells, including stem cells, were collected from corneal limbal tissue. The epithelial cells (1 x105) were co-cultured with MMC-treated 3T3 feeder-layers on temperature-responsive culture dishes for 2 weeks. Cultivated cells were then harvested from the culture dishes by reducing temperature treatment. Some preparations were observed by light and electron microscopy. Autologous transplantation of the cultivated epithelial cells was undertaken to reconstruct the ocular surface in rabbits with total limbal stem cell deficiencies. Results: After 2 weeks in culture, a multilayered corneal epithelial sheet was formed. This was harvested as a transplantable cell sheet simply by reducing the culture temperature to 20 degreeC. Electron microscopy revealed numerous small microvilli in the apical cell membranes of superficial cells, and prominent desmosomes along cell-cell junctions of middle cells. By autologous transplantation, the sheet readily adhered to the underlying corneal stroma, self-stabilizing without any suturing. The rabbit corneal surface was successfully reconstructed and its transparency was maintained during observation periods up to 90 days. Conclusions: Our epithelial cell sheet engineering technique utilizing temperature-responsive culture surfaces permits rapid, adherence of corneal epithelial sheet to the corneal stroma without any carriers or sutures. The results suggest promising clinical capabilities for our bioengineered corneal sheet ideal for transplantation.

(PDF emailed within 1 workday: $29.90)

Accession: 034312533

Download citation: RISBibTeXText


Related references

Functional bioengineered corneal epithelial sheet grafts from corneal stem cells expanded ex vivo on a temperature-responsive cell culture surface. Transplantation 77(3): 379-385, 2004

Autologous transplantation of a bioengineered cultivated oral epithelial cell sheet using a temperature responsive culture surface for ocular surface reconstruction. ARVO Annual Meeting Abstract Search & Program Planner : Abstract No 4695, 2003

Corneal stromal changes following reconstruction by ex vivo expanded limbal epithelial cells in rabbits with total limbal stem cell deficiency. British Journal of Ophthalmology 87(12): 1509-1514, 2003

Corneal surface reconstruction in rabbits with unilateral total limbal deficiency using ex-vivo limbal epithelial stem cells expanded on rabbit amniotic membrane Long term outcomes. IOVS 42(4): S266, March 15, 2001

Ocular Surface Reconstruction with Cultivated Limbal Epithelial Cells in Limbal Stem Cell Deficiency: One-year Follow-up Results. Turk Oftalmoloji Dergisi 42(3): 172-176, 2012

Corneal surface reconstruction by limbal epithelial cells ex vivo expanded on amniotic membrane. IOVS 41(4): S756, March 15, 2000

Human pluripotent stem cell-derived limbal epithelial stem cells on bioengineered matrices for corneal reconstruction. Experimental Eye Research 146: 26-34, 2017

Ex vivo expansion of corneal limbal epithelial/stem cells for corneal surface reconstruction. European Journal of Ophthalmology 13(6): 515-524, 2003

Ocular surface reconstruction using autologous rabbit oral mucosal epithelial sheets fabricated ex vivo on a temperature-responsive culture surface. Investigative Ophthalmology and Visual Science 46(5): 1632-1639, 2005

Ocular surface reconstruction in limbal stem cell insufficiency: transplantation of limbal tissue. Der Ophthalmologe 109(9): 857-862, 2013

S100A expression in normal corneal-limbal epithelial cells and ocular surface squamous cell carcinoma tissue. Molecular Vision 17: 2263-2271, 2012

Ocular surface reconstruction with a tissue-engineered nasal mucosal epithelial cell sheet for the treatment of severe ocular surface diseases. Stem Cells Translational Medicine 4(1): 99-109, 2016

Early Results of Ocular Surface Reconstruction in Unilateral Severe Limbal Stem Cell Deficiency Using Autologous Cultured Limbal and Conjunctival Stem Cells. ARVO Annual Meeting Abstract Search & Program Planner : Abstract No 2992, 2002

Characterization of the corneal surface in limbal stem cell deficiency and after transplantation of cultured allogeneic limbal epithelial cells. Graefe's Archive for Clinical and Experimental Ophthalmology 254(9): 1765-1777, 2017

Amniotic membrane transplantation before limbal stem cell allografts for ocular surface reconstruction in patients with limbal stem cell deficiency. Investigative Ophthalmology & Visual Science 38(4 PART 1-2): S511, 1997