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Human skin equivalent retained functionality after 72 hours

An autologous bilayered human skin equivalent was stored in basal medium at 4°C for at least 72 hours without compromising its functionality for transplantation, according to study results.

Skin samples from six donors in Malaysia were processed within 24 hours from harvest and digested with 0.6% Collagenase Type 1. They were then treated with 2 mL of TrypLE Select to detach fibroblasts from keratinocytes, which were culture-expanded to create sufficient cells to construct the bilayered human skin equivalent, MyDerm. The product was immersed in 2 mL basal F12:DMEM medium, stored at 4°C and evaluated at 0, 24, 48 and 72 hours.

There was no physical change in MyDerm after 72 hours, its cells showed no significant difference in viability (90.5% ± 2.7% to 94.9% ± 1.6%), and they had short population doubling time (58.4 ± 8.7 to 76.9 ± 19 hours). In quantitative gene expression analysis, COL III (0.026 ± 0.008 to 0.087 ± 0.071), CK10 (0.337 ± 0.085 to 0.425 ± 0.098) and CK14 (0.246 ± 0.049 to 0.081 ± 0.027) were comparable during evaluation at 0 and 72 hours, with minimal differences found (P=.436).

“This study demonstrated that MyDerm … showed good viability and population doubling time,” the researchers said. “The long shelf life … enables this skin substitute to be transported to other regions of the country and even to other parts of the world when maintained under good storage conditions.”