Читать книгу Genetic Disorders and the Fetus - Группа авторов - Страница 126

The culture surface has a definite influence on the rate of attachment and proliferation. To attach to the culture surface, AFCs must create their own microenvironment, consisting of glycoproteins, collagen, laminin, and fibronectin, among others (ECM proteins). Fetal bovine serum contains fibronectin and so does human AF at 15–18 weeks of gestation.⁶⁴³ If serum concentrations of less than 10 percent are used (such as in Chang‐type media supplements), the presence of AF in the culture setup might facilitate the coating of fibronectin on plastic surfaces. Chang and Jones⁶⁰⁷ reported optimal cloning and growth when cultures were initiated with equal parts of AF and growth factor‐supplemented medium (including 4 percent fetal bovine serum). Two other studies have shown that precoating the plastic surfaces with ECM improves both cloning and rate of growth of AFCs.^{644, 645} In both laboratories, ECM‐coated dishes were custom made from bovine corneal endothelial cells. The use of such precoated surfaces may be advantageous for cell attachment and cloning if suboptimal media have to be used. It appears impractical for routine use unless the laboratory is prepared to accept the extra expense involved in purchasing precoated dishes. A number of manufacturers offer such “biologic” plastic ware.