Excised Conjunctival Tissues

A variety of automatic voltage clamp devices with special modifications have been extensively utilized in electrophysiological studies of /sc in several ocular tissues including the amphibian corneal epithelium [42] and human fetal retinal pigment epithelium [43, 44], as well as non-ocular tissues like the rat tracheal epithelium [45], A strong temperature dependency and inhibitory effect of serosally instilled ouabain on the rabbit conjunctival /sc are characteristic of active ion transport driven by Na+/K+-ATPases in the conjunctiva [6, 7], 

EXCISE EYEBALL TREATMENT WITH A FROM SOCKET PROTEASE SOLUTION SEPARATE THE CONJUNCTIVA 

CULTURED CELLS ARE PLACED UNDER AIR-INTERFACE CONDITION FROM DAY 4 

Primary Culture Models of Conjunctival Epithelial Cell Layers 

An alternative method for assessing cell layer integrity is through the use of hydrophilic paracellular transport markers (e.g., radiolabeled D-mannitol or fluorescein-Na+), which passively traverse cells by the paracellular route. Small amounts of compound required for in vitro conjunctival cell culture transport experiments make this approach well suited for screening purposes. Relative absorption index of a series of pharmacologically active molecules can be ranked against known markers for the identification of candidates with potential absorption problems, which is a reliable tool to select drug candidates with optimal characteristics. 

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Because mucin and/or cilia systems of AIC cultured epithelial cells may work as a barrier for drug transport, lower Papp values are expected in cell layers cultured in AIC than in LCC methods. However, it was interesting to note that no significant differences in Rapp values were observed between the cell layers cultured with the two methods (Table 9.1). This is in contrast to solute permeabilities reported previously for cell layers cultured with LCC versus AIC methods [76, 80], For example, Yang et al. reported that Rapp of lipophilic solutes (e.g., various /3-blockers) across the primary cultured conjunctival epithelial cell layer are about threefold lower when cultured under AIC than LCC conditions, suggesting that the permeability of AIC cultured cell layers generally better reflects that of the excised tissue than LCC counterparts. Mathias et al. [76] also reported that the permeability of hydrophilic solutes across the primary rabbit tracheal epithelial cell layer cultured under AIC conditions was only half of that observed for cell layers cultured under... 

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