HCEnC-21 and HCEnC-21T cells express genes for ion pumping and typical corneal endothelial markers.

<p>(A) Cell extracts were separated by SDS-PAGE and immunoblotted for N-cadherin and collagen type 8 α2. 21M, HCEnC-21 and HCEnC-21T, but not stromal fibroblasts, synthesized N-cadherin and Col8α2. E: earlier passages <25. L: later passages >45. (B–G) Expression of (B) Na/K ATPase α1, (C) Na/K ATPase α3, (D) carbonic anhydrase 2 (CA2), (E) Na/H⁺ exchanger (NHE1), (F) neuron-specific enolase (NSE), and (G) aquaporin 1 (Aqp1) was detected by real-time PCR. Na/K ATPase α3 and Aqp1 showed increased expression; all other genes were expressed at similar levels relative to 21M primary cells. Stromal fibroblasts expressed significantly less Na/K ATPase α1 and α3, NHE1, and CA2 relative to HCEnC-21 and HCEnC-21T. (H) Monocarboxylate cotransporters (MCT1, -2, and -4), anion exchanger 2 (AE2), carbonic anhydrase 12 (CA12), cystic fibrosis transmembrane conductance regulator (CFTR), soluble adenylyl cyclase 10 (sAC10), and GAPDH were detected by RT PCR. All genes, except for MCT4 and CA12, showed similar expression in HCEnC-21 and HCEnC-21T compared to corneal endothelial tissue. Error bars indicate mean ± SEM. *, p<0.05.