The interaction of cell membranes with the various components of the extracellular matrix (ECM) proteins plays a major role in their regulation and function. The culture of primary tumors or immortalized cell lines on tissue culture dishes in vitro often does not reflect the cellular microenvironment of the cells in vivo. The basement membrane provides cells with the spatial orientation and stability required for the organization and development of the characteristic histology of specific tissues. In addition to its structural function the ECM also plays a role in the migration, chemotaxis and invasion of neoplastic cells during the metastatic cascade.
The Biocoat cell culture inserts are “transwells” that contain a porous membrane at the bottom. This porous membrane is available in different pore sizes - 8µm, 0.45 µm and 0.22 µm. The inserts are available in a range of sizes that fit into 6, 12, 24, 48, 96 and 384-well plates, dividing the wells into the apical chamber (within the insert) and the basolateral chamber (below the insert). Furthermore the inserts are pre-coated with ECM proteins like collagen or Matrigel. I have used the Biocoat inserts in the culture of endothelial cells as well as polarized intestinal epithelial cells (the cell line Caco-2). Using the collagen coated cell culture inserts I have grown Caco-2 cells to study the permeability of therapeutic peptides across the intestinal epithelium. I have also used collagen and Matrigel coated cell culture inserts for in vitro chemotaxis and invasion studies using endothelial cells.
The behavior of cells cultured on the Biocoat inserts is highly consistent and reproducible. The inserts can be stored at 4°C and can be easily rehydrated using warm culture medium. These inserts are compatible with all cell culture media and additives. I have found that the Biocoat cellware closely mimics the functional characteristics of basement membranes in vivo. They are particularly useful for the culture of primary tumors from biopsies.
The Biocoat cell culture inserts are expensive, especially if many samples are being handled at the same time, the experiment with each sample being performed in duplicate or triplicate. They also have a limited shelf life, unlike routine tissue culture inserts. However, they are an excellent tool in the study of cell differentiation, migration and drug permeability studies.
Piyali Dasgupta
Postdoctoral Research Fellow
Department of Pathology
College of Physicians and Surgeons (14-460)
Columbia University, New York