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In Vitro Characterization of Corneal Cells: A Step towards Bioengineered Cornea

E. Sharmila E. Latif 1, Choy Ker Woon2 , Ng Sook Luan2 , Jemaima Che Hamzah2, and Chua Kien Hui2
1 Department of Anatomy, Medical Faculty, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia
2 Department of Anatomy, Physiology and Ophthalmology, Medical Faculty, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia

Abstract—In vitro characterization of cells is an essential step to ascertain the phenotype of the cultured cells prior to the construction of any bioengineered organ. The present study aimed to characterize cultured corneal epithelial cells (CEC) and corneal stromal cells (CSC) with regard to morphology, gene and protein expressions. Corneal cells were isolated and cultured until passage 1 from six New Zealand white strain rabbits’ eyes. The morphology of both cells was examined via phase contrast microscopy. CEC specific differentiation marker, Cytokeratin 3 (CK 3), was analyzed via gene expression and immunocytochemistry. CSC phenotype was analyzed via Aldehyde dehydrogenase (ALDH), Vimentin and alpha-smooth muscle actin (α-SMA) expressions. CEC exhibited polygonal-shaped morphology with the expression of corneal epithelial specific marker, CK 3. Cultured CSC showed mixed phenotypes, both quiescent (ALDH) and active repair phenotypes (Vimentin and α- SMA). The results revealed both cultured CEC and CSC exhibiting suitable phenotype which may be beneficial for application in the construction of bioengineered cornea.

Index Terms—Corneal epithelial cells, Corneal stromal cells, Gene expression, Immunocytochemistry.

Cite: Norzana Abd Ghafar, E. Sharmila E. Latif , Choy Ker Woon , Ng Sook Luan , Jemaima Che Hamzah, and Chua Kien Hui, "In Vitro Characterization of Corneal Cells: A Step towards Bioengineered Cornea," International Journal of Pharma Medicine and Biological Sciences, Vol. 4, No. 1, pp. 29-33, January 2015.
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