Comparing the Effects of Two Cryoprotectant Protocols, Dimethyl-Sulfoxide (DMSO) and Glycerol, on the Recovery Rate of Cultured Keratinocytes on Amniotic Membrane.

This article focuses on evaluating cryopreservation methods for multilayer keratinocyte sheets cultured on decellularized human amniotic membrane (KC sheet-HAM), a tissue-engineered product intended for wound healing applications. The study found that while keratinocytes successfully formed stratified, viable sheets on the amniotic membrane scaffold, cryopreservation using conventional cryoprotectants dimethyl-sulfoxide (DMSO) and glycerol significantly reduced cell viability and disrupted the multilayer structure after thawing. Although some viable cells remained post-thaw, neither cryoprotectant allowed recovery to pre-freeze levels, indicating that current protocols are inadequate for effective biobanking of these tissue constructs. The findings underscore the need to develop improved cryopreservation strategies tailored to maintain the viability and structural integrity of keratinocyte sheets on amniotic membranes for off-the-shelf clinical use.