The pertinence of expression of heat shock proteins (HSPs) to the efficacy of cryopreservation in HELAs.
HELAs (Hela cells, passed cells of human cervical carcinoma) were heat or cold treated (named heat or cold shock) and then resumed normal culture for 2, 4 or 8 hours respectively. The expressions of heat shock protein 70 (HSP70) and 90 (HSP90) of the HELAs were measured by Northern and Western blotting. HELAs after 4-hour culture were exposed to or cryopreserved with different concentration of dimethyl sulfoxide (Me2SO, 2.5%, 5%, 10%, 15% and 20% respectively, V/V). Meanwhile, the HELAs after different culture time (2, 4 and 6 hours of culture) were cryopreserved with 5% Me2SO. After exposure or cryopreservation, the number of live HELAs was counted and the survival rate was calculated. The results showed that heat shock increased the expression of HSP70 and HSP90 of HELAs, while cold shock decreased the expression of the two proteins. When the concentrations of Me2SO were 10%, 15% and 20%, the survival rates of HELAs after exposure to Me2SO or cryopreservation were much lower than those when the concentrations were small. The survival rates of the heat shocked HELAs were significantly higher than those of the cold shocked and control HELAs. After cryopreservation with 5% Me2SO, the survival rate of heat shocked HELAs group with 2 hours culture time was the lowest among all the groups of HELAs with different cultural time. From the results of this study, we conclude that the expressions of HSP70 and HSP90 in HELAs increased significantly after heat shock, while cold shock decreased the expressions of these two proteins. The over-expressions of HSPs in the heat shocked HELAs could protect the cells from both injury caused by potential toxicity of high concentrations of Me2SO and cryoinjury caused by the freeze-thawing/cryopreservation procedure.
Wang
Peitao
P
Department of Thermal Science and Energy Engineering, University of Science and Technology of China, Hefei, China. peit_wang9@ustc.edu
Shu
Zhiquan
Z
He
Liqun
L
Cui
Xiangdong
X
Wang
Yuzhen
Y
Gao
Dayong
D
D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000295:instrumentation / Q000379:methods*; D003451:Cryoprotective Agents / Q000008:administration & dosage; D004121:Dimethyl Sulfoxide / Q000008:administration & dosage; D006367:HeLa Cells / Q000187:drug effects / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000096:biosynthesis*; D006801:Humans; D012333:RNA, Messenger / Q000378:metabolism
D003451:Cryoprotective Agents; D006360:Heat-Shock Proteins; D012333:RNA, Messenger; D004121:Dimethyl Sulfoxide