Improved preparation and preservation of human keratoplasty lenticules.
OBJECTIVE
To improve the preparation of lenticules from human cornea and to obtain their preservation without loss of viable keratocytes.


METHODS
The epithelium was manually removed after bathing the surface of the cornea with a solution of trypsin and EDTA. Lenticules were prepared by microkeratome resection and viable keratocytes were visualized by staining with thiazolyl blue (MTT).


RESULTS
The pretreatment with trypsin-EDTA allowed the removal of the epithelium without damage to the keratocytes and the stroma. When these lenticules were incubated in Optisol-GS for 7 days at 4 degrees C, they showed a limited thickness increase and a preservation of keratocyte viability.


CONCLUSIONS
This procedure allows the preparation of lenticules with viable keratocytes that can be preserved in the cold for at least 1 week.
Salvalaio
Gianni
G
Fondazione Banca degli Occhi del Veneto, Venezia Mestre, Italy.
Fasolo
Adriano
A
Bruni
Alessandro
A
Frigo
Anna Chiara
AC
Favaro
Elisa
E
Ponzin
Diego
D
D002470:Cell Survival / Q000502:physiology; D002809:Chondroitin Sulfates; D045424:Complex Mixtures; D003315:Cornea; D003319:Corneal Stroma / Q000166:cytology*; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000627:therapeutic use; D016895:Culture Media, Serum-Free; D003646:Debridement; D003911:Dextrans; D005347:Fibroblasts / Q000166:cytology*; D005839:Gentamicins; D006801:Humans; D015948:Keratoplasty, Penetrating; D009926:Organ Preservation / Q000379:methods*; D013194:Staining and Labeling / Q000379:methods
D045424:Complex Mixtures; D003451:Cryoprotective Agents; D016895:Culture Media, Serum-Free; D003911:Dextrans; D005839:Gentamicins; D002809:Chondroitin Sulfates; C075736:Optisol