miR profiles in fresh corneal endothelial tissues. The tissues were a gift from SightLife, Inc. It should be noted that no diagnosis was performed to determine whether or not the donors of the endothelium tissues with gutatta were afflicted with Fuchs' endothelial corneal dystrophy (FECD). (
A) Images of fresh corneal endothelial tissues with guttata (donor age: 73 years, donor ECD: 378 cells/mm
2; donor age: 73 years, donor ECD: 1552 cells/mm
2; and donor age: 51 years, donor ECD: 2457 cells/mm
2). (
B) Scatter plots of cHCEC miR expression profiles. The Toray 3D-Gene forms of Ver. 19/20 were converted to Ver. 17, and the values and the
upper/lower of 2-fold lines are as explained for
Figure 1. The
upper plot shows the comparison of tissues with normal-level ECD (donor age: 3 days, donor ECD: 7470 cells/mm
2; donor age: 7 months, donor ECD: 5070 cells/mm
2; donor age: 12 months, donor ECD: 4551 cells/mm
2; donor age: 12 years, donor ECD: 3987 cells/mm
2; donor age: 20 years, donor ECD: 3812 cells/mm
2; donor age: 63 years, donor ECD: 2598 cells/mm
2, Ver. 17) (donor age: 66 years, donor ECD: 2918 cells/mm
2, Ver. 20) with tissues with lowered ECD of 378 cells/mm
2 (donor ECD: 378, donor age: 73 years, Ver. 19). The
lower plot shows the comparison among tissues with guttata but different in ECD. The tissues were those with an ECD of 378 cells/mm
2 and averaged 2318 cells/mm
2 (donor age: 50 years, donor ECD: 2187 cells/mm
2, donor age: 51 years, donor ECD: 2379 cells/mm
2, Ver. 17). (
C) The normalized value of miR378a-5p and 378f in corneal endothelial and epithelial tissues is depicted. The miR378f expression level was dramatically decreased in tissues with lowered ECD. (
D) The normalized value of miR146a-5p and 146b-5p in corneal endothelial/epithelial tissues, cHCECs, and supernatants of cHCECs are shown. CST+ means cHCECs with morphologically abnormal cells differing from CD44
−/+. The miR146b-5p expression levels were uniformly upregulated during cultures. (
E) Expression levels of miR-378a-3p, -378e, and -378f were evaluated by qRT-PCR. The miRs were extracted from fresh HCE tissues distinct in ages and ECDs (donor age: 54 years, donor ECD: 3107 cells/mm
2, donor age: 72 years, donor ECD: 1411 cells/mm
2 [guttata+], donor age: 60 years, donor ECD: 795 cells/mm
2 [guttata+]).