Cell Sheet(CS) Engineering is based on the retrieval of cultured cells as ECM-rich sheets that can be used therapeutically, as demonstrated in ongoing clinical trials. CS fabrication, while simple, requires cell culture facilities. For its efficiency it is essential that adequate cell function from the fabrication site to the bedside is maintained. We tested the ability of two commercially available compounds to preserve the viability of CS of human adipose stem cells (hASC) at 4ºC. hASC were cultured to hyperconfluence in basal medium to produce CS, and then culture medium was either replaced by a solution of Hypothermosol®(HTS) or supplemented with Rokepie®(RP) and cells maintained at 4ºC for 3 and 7 days. Basal medium controls(BMC) at 4ºC and 37ºC, together with non-confluent cultures, were also established. At each time point, cell viability, caspase activity and differentiation potential into adipogenic and osteogenic lineages were assessed. After 3 days, a sharp decrease in cell viability in 4ºC BMC was noticed, especially in non-confluent cultures. HTS and RP managed to keep viability values close to the 37ºC BMC. At day 7, few viable cells were detected in the 4ºC BMC, while none were found in their non-confluent counterparts. HTS- and RP-supplemented CS presented significantly more viable cells than non-confluent conditions. Lack of activation of caspase 3 suggests cell necrosis is the main process behind cell loss. Both HTS and RP where equally able to preserve hASC ability to differentiate into adipogenic lineage, whereas, RP demonstrated higher efficiency in preserving hASC ability to differentiate into osteogenic lineage. Summarizing, both HTS and RP demonstrated excellent hypothermic preservation of cells especially up to 3 days. Acknowledgements: The author acknowledges the Project PD/59/2013, IF/00347/2015, SFRH/BPD/96611/2013. NORTE-01-0145-FEDER-000021, Gene2Skin Project (H2020-TWINN-2015-692221)