In this study, we had dealt with the improved cultures of cHCECs nearly without conspicuous EMT, although it is difficult to discriminate the presence of a minuscule amount of EMT among heterogeneous cHCECs. At first, we compared the profiles of miRs between a CD44
− SP (effector cell, 66-P4) and 55-P5 with senescent-like CST (
Fig. 1A). The senescent-like CST was confirmed on the grounds of elevated SA-β-Gal staining and their phenotypes with CD44
++, senescent-type, nonhexagonal cells forming island-like clusters, distinct from CD44
− mature, differentiated, hexagonal, cobblestone-shape cells. The cHCECs with the senescent-like CST exhibited cell areas that were nearly twice as large as the latter mature cells.
20 The scatter plot depicted in
Figure 1B indicates the upregulation of several miRs in 55-P5 compared to 66-P4. The upregulated miRs are summarized in
Figure 1C. Of note, some of the miRs present in the control medium were greatly decreased in CS during the 2-day cultures of 66-P4, although no decreases in CS were observed in the cultures of 55-P5. These miRs were miRs-1246, 1273g-3p, and 4732-5p. Simultaneously, other groups of miRs, such as miRs-1273f, 1285-3p, 1972, and 221-3p, were significantly increased in CS of the 55-P5 culture than in the control medium. These two typical changes of miRs in CS also were confirmed by the comparison of 66-P4 with other cHCECs (i.e., 72-P3 and 69-P3) with evident morphologic CST.