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Culturing hepatocytes and other differentiated cells

1984; Lippincott Williams & Wilkins; Volume: 4; Issue: 3 Linguagem: Inglês

10.1002/hep.1840040332

1527-3350

Lola M. Reid, Douglas M. Jefferson,

Pluripotent Stem Cells Research

Establishment of normal adult cells in culture is exceedingly difficult.In recent years, culture conditions have been improved to permit maintenance of some normal cell types for weeks or months in culture.In this review, we will compare methodologies for culturing normal cells, emphasizing studies on liver and providing findings with other cell types for comparison.In all systems, the basic protocol incorporates tissue culture plastics, basal media (media which are defined for amino acids, salts, sugars and other nutrients) and supplementation of either serum or some other biological fluid (e.g., lymph, embryo extracts, etc.) or defined and purified factors.We will also review the use of co-cultures, a technique by which two or more cell types are cultured together resulting in cell behavior and physiological responses which would not occur if the cell types were cultured alone.The impasse for culturing normal cells or even differentiated neoplastic cells, has been overcome by use of co-cultures.Only in recent years has co-culture been supplanted, in a few systems, with more defined cell culture conditions.We will also present current findings with co-cultures and defined conditions which replaced co-cultures in some systems.Co-culturing was first described by Puck and Marcus in 1955 in studies with HeLa cells (1).Throughout the history of studying cultured cells, co-culture has been a frequent approach to elicit more differentiated responses even in cells long adapted to classical cell culture conditions.Moreover, one of the most successful methods for maintaining functional primary cultures is to use irradiated or mitomycin-C-treated "feeder layers" of another cell type (often fibroblasts), which are thought to "feed" the primary cells with essential nutrients or other factors (1-3).Success in co-cultures, especially with primary cultures, is predicated on an important principle of cel-