부산대학교 도서관

Safety concerns associated with foetal bovine serum (FBS) have restricted its translation into clinics. We hypothesised that platelet lysate (PL) can be utilised as a safe alternative to produce serum-free 3D-engineered skin. PL supported a short-term expansion of fibroblasts, with negligible replication-induced senescence and directed epidermal stratification. PL-expanded fibroblasts were phenotypically separated into three subpopulations of CD90.sup.+FAP.sup.+, CD90.sup.+FAP.sup.- and CD90.sup.-FAP.sup.+, based on CD90 (reticular marker) and FAP (papillary marker) expression profile. PL drove the expansion of the intermediate CD90.sup.+ FAP.sup.+ subpopulation in expense of reticular CD90.sup.+FAP.sup.-, which may be less fibrotic once grafted. The 3D-engineered skin cultured in PL was analysed by immunofluorescence using specific markers. Detection of ColIV and LMN-511 confirmed basement membrane. K10 confirmed near native differentiation pattern of neo-epidermis. CD29- and K5-positive interfollicular stem cells were also sustained. Transmission and scanning electron microscopies detailed the ultrastructure of the neo-dermis and neo-epidermis. To elucidate the underlying mechanism of the effect of PL on skin maturation, growth factor contents in PL were measured, and TGF-[beta]1 was identified as one of the most abundant. TGF-[beta]1 neutralising antibody reduced the number of Ki67-positive proliferative cells, suggesting TGF-[beta]1 plays a role in skin maturation. Moreover, the 3D-engineered skin was exposed to lucifer yellow on days 1, 3 and 5. Penetration of lucifer yellow into the skin was used as a semi-quantitative measure of improved barrier function over time. Our findings support the concept of PL as a safe and effective serum alternative for bioengineering skin for cell therapies.
Author(s): I. Banakh [sup.1] [sup.2], Md. M. Rahman [sup.1] [sup.2], C. L. Arellano [sup.1] [sup.2], D. C. Marks [sup.3] [sup.4], S. Mukherjee [sup.5], C. E. Gargett [sup.5], H. Cleland [sup.1] [...]