Figure 2 | Scientific Reports

Figure 2

From: Regulatory Compliant Tissue-Engineered Human Corneal Endothelial Grafts Restore Corneal Function of Rabbits with Bullous Keratopathy

Figure 2

Comparative assessments of cell culture reagents. (A) Human CEnCs were seeded onto wells that were either pre-coated for FNC coating, human Collagen IV (blue), hLaminin-511 (green) or hLaminin-521 (yellow), or non-coated (white) as a negative control and analyzed for up to 48 hours using the xCELLigence system. Normalized impedance readings of CEnCs were extracted at 8, 24, and 48 hours. Cell index was normalized to 1.0 (red line) to cells seeded on FNC-coated wells for each of the time points. Statistical significance were detected between the impedance index of CEnCs seeded onto control vs FNC-coat wells (n = 4, *p < 0.05) at 8 hours, as well as between CEnCs seeded onto hLaminin-511 vs FNC-coated wells (n = 4, *p < 0.05). (B) Representative images showing the morphology of 3 confluent cultures of donor-matched human CEnCs grown in regular FBS, Equafetal, or commercially obtained HS. Scale bar: 100 µm. (C) Flow cytometric dot plots of overall percentage live cells (red box), as revealed by the Annexin V detection kit following cryo-preservation in current freezing solution of M5-Endo containing 10% DMSO, CryoScarless, and FREEZEStem. Insert within each condition showed morphology of cryo-preserved CEnCs that were revived directly into a high density seeding following cryo-preservation in its respective freezing medium. Scale bar: 100 µm.

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