The Role of Fibroblasts in Tissue Engineering and Regeneration

T. Wong; J.A. McGrath; H. Navsaria

Disclosures

The British Journal of Dermatology. 2007;156(6):1149-1155. 

In This Article

Culturing Fibroblasts

The development of techniques for culturing fibroblasts was long established prior to the discovery made in 1975 by Rheinwald and Green for culturing and expanding keratinocytes, which require growth-arrested murine 3T3 fibroblast cells to support their proliferation.[3] Dermal fibroblasts can be extracted from skin biopsies either through enzymatic degradation or by explant culture, which is particularly useful for obtaining cells from smaller specimens. The medium used for culturing fibroblasts is usually supplemented with fetal calf serum which previously raised concerns regarding transmission of bovine spongiform encephalopathy (BSE). However, the serum that is currently used is obtained only from BSE-free countries.[4]

Growth parameters and the characteristics of fibroblasts in culture will be influenced by passage number, age of the donor, subtype of fibroblast (reticular or papillary dermis) being cultured and anatomical site. Older donor skin fibroblasts compared with younger skin fibroblasts tend to migrate more slowly, reach cell culture senescence earlier and have a prolonged cell population doubling time.[5] In addition, elderly donor fibroblasts are less responsive to growth factors such as platelet-derived growth factor, epidermal growth factor, dexamethasone, insulin and transferrin in vitro.[6]

Other factors that influence fibroblast behaviour in culture include vitamins, such as vitamin C, and antioxidants, including coenzyme Q10. For example, in the presence of vitamin C 100 µmol L-1, fibroblasts produce twofold more collagen than fibroblasts cultured without, a response that is independent of the age of the fibroblasts.[6] Likewise, coenzyme Q10 supports wound healing by increasing cell proliferation and fibroblast mobility when compared with fibroblasts cultured without this antioxidant.[7]

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