A modified culture strategy of human keratinocytes to shorten the primary culture time.

Shortening time from biopsy to graft preparation is crucial for the coverage of extensive burn wound in major burn patients. This study was aimed to shorten the culture time of keratinocytes by means of one modified cell culture method. Three factors (temperature of dispase, area of skin, and age of patient) that might have impacts on cell behavior were evaluated. The results showed that cells isolated with dispase at 37°C exhibited an improved cell activity than being isolated at 4°C. The time that cells reached subconfluence with various skin areas was in order of 3 cm(2) < 2 cm(2) < 1 cm(2) (120.07 ± 5.03 h, 141.33 ± 4.16 h, and 193.33 ± 6.11 h, respectively). Furthermore, compared with the young and middle-aged groups, proliferation of keratinocytes isolated from children skin tissue was better. These results indicated that 3 cm(2) skin biopsy from a young patient, isolated with dispase at 37°C, was a promising strategy to shorten the cultivation of keratinocytes.

The roles of autophagy and apoptosis in burn wound progression in rats.

Understanding the role of cell death in burn wound progression is crucial for giving appropriate diagnoses and designing therapy regimens for burn patients. A well-described and reliable "comb burns model" was employed to evaluate the roles of autophagy and apoptosis in burn wound progression at 2 h, 6 h, 12 h, 24 h, and 48 h post-burn in a rat model. Immunohistochemistry (IHC) results showed that autophagy was detectable in hair follicle epithelium at 2 h post-burn, peaked at 12 h post-burn, then declined. Conversely, apoptosis was mainly located in the stratum epidermis and took place at low levels until 6 h post-burn, at which point it slowly increased. Bcl-2 and Bax, which are regulators of both processes, showed protein expression level patterns that were consistent with the IHC results. This study of autophagy in burn wound tissue progression represents a conceptual expansion of cell death in burn wounds. Based on these results, we suggest that different treatments should be performed on a specific post-burn time course depending on the most prevalent type of cell death occurring at that time.