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Introduction
Co-culture describes various techniques where different cellpopulations are cultivated in close proximity in the same cellculture environment. The applications of co-cultures are multifacettedand include:

stimulation and maintenance of cell function and differentiation,
cultivation of embryonic stem cells on feeder cells,
applications in reproductive medicine (e.g. autologous endometrial co-culture),
investigation of immune cell interactions,
investigation of paracrine mesenchymal-epithelial interactions,
restoration of heterocellular functions in vitro (e.g. blood-brain-barrier).

Direct co-culture can be performed in nearly all cell culturedishes, for instance by layering two cell types one on top ofthe other. In contrast, indirect co-culture takes advantage ofcell culture inserts with porous membranes, to keep the cocultivatedcell populations separated (Fig. 1).

Co-cultures with cell culture inserts have been widely used tostudy mesenchymal-epithelial interactions during normal andtumoral development (Hofland et al., 1995; Gache et al.,1998). Here, such a co-culture model has been establishedusing MCF7 mammary carcinoma cells, human fibroblastsand ThinCert^{/sup> cell culture inserts. These experiments illustratethe excellent suitability of ThinCert/sup> cell culture insertsfor co-culture applications.}

The following protocol presents technical details for co-cultureand may be easily adapted to match individual requirementsand research interests other than paracrine growth regulation.

Material

Methods
Seeding of fibroblasts onto the underside of themembrane and co-culture
24 well ThinCert^{/sup> cell culture inserts with translucentmembranes and 0.4 pores were inverted and placed in a12 well plate (Fig. 2/1). The well bottom was humidified with100 sterile water (Fig. 2/2). 60 of a cell suspensioncontaining 0 (control); 83,000; 167,000 or 418,000 humanjuvenile foreskin fibroblasts per ml DMEM medium(supplemented with 10% FCS, 4 mM L-alanyl-glutamine) waspipetted into the inner circle of the underside of the membrane(Fig. 2/3, 2/4).}

Related article:

• Co-culture in ThinCert cell culture inserts2008-04-09 07:50:39