Publications (3)10.95 Total impact
Article: Development of defined media for the serum-free expansion of primary keratinocytes and human embryonic stem cells.[show abstract] [hide abstract]
ABSTRACT: Primary keratinocyte (Kc) cells and human embryonic stem (hES) cells are routinely propagated on a mouse fibroblast feeder layer in media containing fetal bovine serum or other nondefined factors. One disadvantage of using these nondefined factors is that they may inadvertently contaminate the culture system with infectious agents; thus, there remains a need to develop safe culture conditions free from poorly defined and/or animal products. Our laboratory has discovered that growth factors (GFs) and vitronectin (VN) can bind to each other resulting in synergistic short-term functional effects in several cell types. The aim of the current study was to determine whether primary Kc and hES cells can be established and serially propagated serum-free using medium containing VN, insulin-like growth factor-I, and insulin-like growth factor binding protein-3 (VN:GF). Here we demonstrate that primary Kc cells can be isolated, established, serially propagated, and re-form an epidermal layer using the VN:GF combination. Additionally, cell proliferation studies indicate that the Kcs proliferate using the VN:GF combination at a rate comparable to cells grown using serum. Similarly, we verified that this VN:GF combination could be employed for the serial propagation of hES cells. Importantly, both the Kc and hES cells retain their undifferentiated phenotype when cultured using the VN:GF combinations as a serum-free medium for up to 4 passages for Kc and at least 10 passages for hES cells as indicated by the expression of a range of cell surface markers. This study demonstrates that the novel, fully defined VN:GF medium is a viable alternative to media containing serum and highlights the potential of this technology for generating therapeutically viable cells and tissues.Tissue Engineering Part C Methods 08/2008; 14(3):221-32. · 4.64 Impact Factor
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ABSTRACT: Topical administration of growth factors has displayed some potential in wound healing, but variable efficacy, high doses, and costs have hampered their implementation. Moreover, this approach ignores the fact that wound repair is driven by interactions between multiple growth factors and extracellular matrix (ECM) proteins. We report herein that complexes comprising IGF and IGF-binding proteins bound to the ECM protein vitronectin (VN) significantly enhance cellular functions relevant to wound repair in human skin keratinocytes in two- and three-dimensional in vitro cell models and are active, even in the presence of wound fluid. Moreover, these responses require activation of both the IGF receptor and the VN-binding alpha(v) integrins. Further, we assessed the complexes as a topical agent in the treatment of deep dermal partial thickness burns in a porcine model. This pilot study revealed that the complexes may hold promise as a wound healing therapy. Critically, the significant responses observed in vitro and the encouraging preliminary data in vivo were obtained with nanogram doses of growth factors. This suggests that coupling delivery of growth factors to ECM proteins such as VN may ultimately prove to be a more effective strategy for developing a wound healing therapy.Journal of Investigative Dermatology 07/2008; 128(6):1535-44. · 6.31 Impact Factor
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ABSTRACT: Chronic ulcers are a major problem affecting a significant number of people around the world. The condition is difficult to heal and often leads to amputation. Hyperbaric oxygen (HBO) has been used clinically for the treatment of chronic ulcers and positive outcomes have been reported. However, owing to the lack of large randomised controlled trials and some conflicting data, controversy regarding the effectiveness of HBO in chronic wound healing persists. Besides randomised controlled clinical trials, in vitro studies hold promise in providing further insight into the role of HBO in wound healing and in aiding the establishment of a scientific foundation upon which more rational and efficacious HBO therapeutic regimes may be developed. The present article provides an overview of the available in vitro data on HBO with regards to wound healing. In particular, it focuses on experimental design issues and future opportunities using human skin equivalent models to study HBO-mediated wound healing.