[Effects of mouse adipose-derived stem cell conditioned medium on the apoptosis of keratinocytes induced by thermal injury in vitro].

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Abstract (original English)

To investigate the effects of mouse adipose-derived stem cell conditioned medium (ADSC-CM) on apoptosis of keratinocytes (human epithelial cell line HaCaT) induced by thermal injury in vitro. (1) Adipose-derived stem cells (ADSCs) from inguinal adipose tissue of 5 healthy BALB/c mice were isolated, cultured, and purified by collagenase digestion in vitro. The 3rd passage of cells were collected for morphologic observation, detection of expressions of surface markers CD31, CD34, CD45, CD90, and CD105 with flow cytometer, and identification of adipogenic and osteogenic differentiation. (2) HaCaT cells were incubated in water at 51.5 °C for 35 seconds to reproduce thermal injury model, and then the apoptosis rate was detected immediately after injury by flow cytometer. (3) Thermally injured HaCaT cells were divided into routine culture group (RC, cultured with DMEM containing 10% FBS), serum-free group (cultured with serum-free DMEM), 50%ADSC-CM group (cultured with DMEM containing 50%ADSC-CM), and 100%ADSC-CM group (cultured with 100%ADSC-CM) according to the random number table. After 24 hours, apoptosis of HaCaT cells was observed by acridine orange-ethidium bromide (AO-EB) staining; apoptotic rate was determined by flow cytometer; the mRNA and protein levels of Bcl-2 and caspase-3 were respectively determined by real-time fluorescent quantitative RT-PCR technique and Western b

Evidence level

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

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