Cryopreservation of tissue-engineered dermal replacement in Me2SO: Toxicity study and effects of concentration and cooling rates on cell viability.
Cryopreservation of tissue-engineered human dermal replacement plays an important role in skin tissue engineering and skin banking. With the inspection of electronic scanning microscope and viability evaluation by Trypan Blue staining assay and the tetrazolium salt, MTT (3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay, this study investigated the toxicity of Me(2)SO to dermal fibroblasts and effects of cryoprotectant concentration and cooling rate on the viability of dermal replacement. The results demonstrated that the Me(2)SO toxicity to fibroblasts was affected by the exposure time, temperature, and concentration. Furthermore adding cryoprotectant solution at low temperature of 4 degrees C significantly reduced the toxic effect on the tissue-engineered dermal equivalent. An optimal cryopreservation protocol consisting of cooling rate at 1 degrees Cmin(-1) in 10% (V/V) Me(2)SO was derived, with the viability of studied dermal equivalent treated by this protocol being 75% of that of fresh control. The micrograph obtained by electronic scanning microscope also confirmed this result.
Wang
Xin
X
Institute of Cryomedicine, Shanghai University of Science and Technology, Shanghai 200093, China.
Hua
Tse-Chao
TC
Sun
Da-Wen
DW
Liu
Baolin
B
Yang
Guanghui
G
Cao
Yilin
Y
D002470:Cell Survival; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000633:toxicity*; D004121:Dimethyl Sulfoxide / Q000633:toxicity*; D005347:Fibroblasts / Q000187:drug effects* / Q000648:ultrastructure; D006801:Humans; D008855:Microscopy, Electron, Scanning; D013997:Time Factors; D023822:Tissue Engineering
D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide