[Reproduction of a model of heat injured keratinocyte in vitro and observation on its apoptosis rate].
OBJECTIVE
To reproduce a model of heat injured KC in vitro and explore its apoptosis rate of KC due to heat injury at different temperature.


METHODS
Human KCs were cultured in vitro, and they were incubated at 37, 41, 43, 45, 48, and 51 degrees C respectively for 10 mins in water bath. Trypan blue staining and Hoechst 33258 fluorescence staining were used respectively to determine necrosis and apoptosis of KC. Rates of apoptosis and necrosis of KC were analyzed quantitatively by flow cytometer. The proliferation activity of KC after heat injury was detected by MTT test.


RESULTS
The results of trypan blue staining, Hoechst 33258 fluorescence staining, and flow cytometer demonstrated that number of apoptotic and necrotic KC increased gradually along with a rise of water bath temperature. The rates of apoptosis and necrosis of KC were respectively (12.3 +/- 3.2)% and (14.1 +/- 1.6)% at 45 degrees C, (27.7 +/- 5.1)% and (58.0 +/- 4.2)% at 48 degrees C. Rate of KC necrosis reached up to (83.0 +/- 5.3)% at 51 degrees C. Inhibition of KC growth reached a stationary phase when the injurious temperature reached 45 degrees C as observed with MTT test.


CONCLUSIONS
Heat injury can induce apoptosis and growth inhibition of KC in vitro. Incubating KC at 45 degrees C for 10 mins is a good condition to reproduce a model of heat injured KC in vitro. This model may be used to study the biological character and apoptosis of KC after burn injury.
Bai
Xiao-Zhi
XZ
Burn Center of PLA, Xijing Hospital, the Fourth Military Medical University, Xi'an 710032, China.
Lü
Gen-Fa
GF
Xie
Song-Tao
ST
Hu
Da-Hai
DH
Zhu
Xiong-Xiang
XX
Tang
Chao-Wu
CW
D017209:Apoptosis; D002056:Burns; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D005434:Flow Cytometry; D006358:Hot Temperature; D006801:Humans; D015603:Keratinocytes / Q000166:cytology*