Cell line/type | Skin explants |
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Species | Human |
Animal free | Yes |
Product | Williams’ E medium Kleszczyński et al. 2012 |
Kleszczyński, K., & Fischer, T. W. (2012). Development of a short-term human full-thickness skin organ culture model in vitro under serum-free conditions. Archives of dermatological research, 304(7), 579-587. This report aims to validate a serum-free human full-thickness skin organ culture model with regard to dependency of skin viability on culture duration and location of skin cross-section (1-5) from the outer (section 1) to the inner side (section 5) of a skin specimen (5 x 10 mm). Cultured skin was analyzed in time-dependent manner for structural damage (H&E staining) and 'balance' between proliferation (Ki67) and apoptosis [cleavage of caspase-3, lactate dehydrogenase (LDH), TUNEL]. First structural disturbances were observed at 48 h (section 3; middle part), increasing with prolonged culture time. Cleavage of caspase-3 and appearance of apoptotic [TUNEL(+)] cells showed significant increase at 72 h in sections 4 and 5, respectively. This correlated to increasing LDH release. Parallel analysis of proliferating [Ki67(+)] cells revealed simultaneous down-regulation within the first 48 h reaching complete absence of Ki67(+) cells at 72 h. These data define an accurate, standardized and robust serum-free short-term ex vivo human full-thickness skin model which is suitable for experimental studies of up to 48 or 72 h in vitro. Skin samples were cultured in the presence of 2 ml serum-free Williams’ E medium. The formulation of serum-free Williams’ E medium can be found online: https://www.sigmaaldrich.com/life-science/cell-culture/learning-center/media-formulations/williams.html. https://link.springer.com/content/pdf/10.1007%2Fs00403-012-1239-z.pdf |
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Source | Literature - own formulation |
Chemically defined > Yes | Yes |
Contains phenol red > Unspecified | Optional |
Antibiotics free > Yes | Yes |