Mixture of Fibroblasts and Adipose Tissue-Derived Stem Cells Can Improve Epidermal Morphogenesis of Tissue-Engineered Skin

Abstract

Many studies demonstrate that the type of adjacent mesenchymal cells can affect epidermal morphogenesis of bilayered tissue-engineered skin. However, whether a mixture of different mesenchymal cell types can improve epidermal morphogenesis of bioengineered skin remains unknown. In this study, keratinocytes, dermal fibroblasts and adipose tissue-derived stem cells (ADSCs) were isolated and purified from human skin and subcutaneous fat. Conditioned medium generated from a mixture of dermal fibroblasts and ADSCs at the ratio of 1:1 was superior to that from fibroblasts or ADSCs alone in promoting keratinocyte proliferation, as indicated by MTT assay. Furthermore, ELISA results showed that the cytokine levels of human hepatocyte growth factor and keratinocyte growth factor (also known as FGF7) in the mixed fibroblasts/ADSC group were higher than those in the ADSC or dermal fibroblasts group. To examine the potential roles of mixed fibroblasts and ADSCs on epidermal morphogenesis, a three-dimensional tissue engineered skin system was applied. Histological analyses demonstrated that keratinocytes proliferated extensively over the mixture of fibroblasts and ADSCs, and formed a thick epidermal layer with well-differentiated structures. Keratin 10 (epidermal differentiation marker) was expressed in the suprabasal layer of bilayered tissue-engineered skin in the mixed fibroblasts and ADSCs group. Desmosomes and hemidesmosomes were detected in the newly formed epidermis by transmission electron microscopy analysis. Together, these findings revealed for the first time that a mixture of fibroblasts and ADSCs in bilayered tissue-engineered skin can improve epidermal morphogenesis.

Bone tissue engineering by using a combination of polymer/Bioglass composites with human adipose-derived stem cells

“…ADSCs used in this study were purchased commercially and showed typical mesenchymal stem cell characteristics, confirmed using multiple differentiation and flow cytometry analysis. The results were consistent with previous publications (Lu et al 2012;Zuk et al 2002).…”

“…Mesenchymal stem cell surface markers were confirmed for cultured ADSCs using fluorescence-activated cell sorting analysis for CD29 and CD44 (eBioscience, San Diego, CA, USA) according to the manufacturer's protocol. The induction of osteogenesis and adipogenesis were as previously reported (Lu et al 2012).…”

Adipose-Derived Stem Cells in Tissue Regeneration: A Review

“…In support of paracrine action, keratinocyte and/or fibroblast proliferation, together with collagen transcription/synthesis by fibroblasts, can be enhanced using ASCconditioned medium or coculture systems [289][290][291]. Conditioned medium made from a mixture of ASCs and dermal fibroblasts can also increase keratinocyte proliferation and migration [292,293] and increased levels of HGF and KGF can be found in the media, suggesting a role for these factors [293]. Similar to this, stromal layers derived from human ASCs can induce the proliferation and expression of appropriate epidermal keratins when overlaid with keratinocytes, leading the authors to conclude that the ASC can be used to create a biomimetic stroma capable of stimulating epidermal development [136].…”

Morphogenesis of Rete Ridges in Human Oral Mucosa: A Pioneering Morphological and Immunohistochemical Study

“…ERK1/2 activation induced by mechanical force has been reported [Matsubayashi et al, 2004;Reichelt, 2007;Quaglino et al, 2009;Perera et al, 2010;Zhang et al, 2012]. However, whether mechanical stress can induce ERK1/2 activation in oral epithelial cells is unclear.…”

“…Recent studies also showed that stromal cells and their matrices can influence upper epidermis development [Carriel et al, 2011;Lu et al, 2012]. Hence, RR elongation may result from the combination of all of the above mentioned aspects.…”