Differential Effects of Tissue Culture Coating Substrates on Prostate Cancer Cell Adherence, Morphology and Behavior

Abstract

Weak cell-surface adhesion of cell lines to tissue culture surfaces is a common problem and presents technical limitations to the design of experiments. To overcome this problem, various surface coating protocols have been developed. However, a comparative and precise real-time measurement of their impact on cell behavior has not been conducted. The prostate cancer cell line LNCaP, derived from a patient lymph node metastasis, is a commonly used model system in prostate cancer research. However, the cells’ characteristically weak attachment to the surface of tissue culture vessels and cover slips has impeded their manipulation and analysis and use in high throughput screening. To improve the adherence of LNCaP cells to the culture surface, we compared different coating reagents (poly-l-lysine, poly-l-ornithine, collagen type IV, fibronectin, and laminin) and culturing conditions and analyzed their impact on cell proliferation, adhesion, morphology, mobility and gene expression using real-time technologies. The results showed that fibronectin, poly-l-lysine and poly-l-ornithine improved LNCaP cells adherence and provoked cell morphology alterations, such as increase of nuclear and cellular area. These coating reagents also induced a higher expression of F-actin and reduced cell mobility. In contrast, laminin and collagen type IV did not improve adherence but promoted cell aggregation and affected cell morphology. Cells cultured in the presence of laminin displayed higher mobility than control cells. All the coating conditions significantly affected cell viability; however, they did not affect the expression of androgen receptor-regulated genes. Our comparative findings provide important insight for the selection of the ideal coating reagent and culture conditions for the cancer cell lines with respect to their effect on proliferation rate, attachment, morphology, migration, transcriptional response and cellular cytoskeleton arrangement.

Differential Effects of Coating Materials on Viability and Migration of Schwann Cells

“…Notably, a study using a prostate adenocarcinoma cell line together with collagen I, fibronectin, laminin, poly- l -lysine, and poly- l -ornithine coating showed conflicting results [32]. In that experiment, a migration-stimulatory effect was only demonstrated for laminin whereas all other coatings were inferior to the polystyrene controls.…”

The use of collagen-based scaffolds to simulate prostate cancer bone metastases with potential for evaluating delivery of nanoparticulate gene therapeutics

“…LNCaP cells are known to be poorly adherent to tissue culture plates and are thus frequently pre-coated with substrates to improve their adherence [47]. However, no such treatment was applied to scaffolds and yet cells adhered to and proliferated on the scaffolds, although to a lesser extent relative to 2D cell culture ( Fig.…”

Silk scaffolds connected with different naturally occurring biomaterials for prostate cancer cell cultivation in 3D

“…After characterizing the various SF scaffolds, they were used to culture the epithelial prostate cancer cell line LNCaP and analyze the influence of the biochemical and physical properties on proliferation as well as spheroid formation. It is known that LNCaP cells are poorly adherent to tissue culture plates and thus a pre‐coating with substrates (eg poly‐L‐lysine) is favorable to improve it . The pure SF scaffold was blended with different natural biomaterials to influence positive attachment and proliferation behavior of the cells.…”