Tissue culture polystyrene

In a separate study [88], we synthesized EGF fused to a polystyrene-binding peptide [101] (EGF-PSt) that could be immobilized on the surface of a tissue culture polystyrene dish. This surface also permitted efficient expansion of NSCs. Thus, EGF-PSt can be used to produce large quantities of pure NSCs in standard laboratories. 

Fig. 2. Effect of serum concentration on the attachment and spreading of BHK-21 cells onto TCP2 surface. BHK-21 cells were seeded in media containing the indicated concentrations of intact serum (open squares), Fn-depleted serum (triangles). Vn-depleted serum (circles), or serum-free medium alone (the single closed square) and the attachment panel (A) and spreading panel (B) of the cells were determined after 90 min culture on TCP (panel (A, B)) Mean SEM. (Reproduced from J. Biomed. Mater. Res. [Ref. 11 Role of serum vitronection and fibronectin in adhesion of fibroblasts following seeding onto tissue culture polystyrene] through the courtesy of John Wiley Sons, Inc.) BHK-21 Fibroblast cell lines from Baby Hamster Kidney 2 Similar results on Primaria are also presented in [Ref 11]...

Steele JG, Dalton BA, Johnson G et al (1993) Polystyrene chemistry affects vitronectin activity - an explanation for cell attachment to tissue-culture polystyrene but not to unmodified polystyrene. J Biomed Mater Res 27(7) 927-940... 

Select proteins that mediate adhesion of specific anchorage-dependent cells (such as osteoblasts, fibroblasts, and endothelial cells) on substrate surfaces have been identified (Underwood and Bennett, 1989 Thomas et al., 1997 Ayad et al, 1994). For example, adsorption of fibronectin and vitronectin on tissue-culture polystryene subsequently enhanced osteoblast, fibroblast, and endothelial cell adhesion (Underwood and Bennett, 1989). More importantly, fibronectin and vitronectin adsorption on borosilicate glass, in a competitive environment, maximized fibroblast and osteoblast adhesion, respectively (Thomas et al., 1997). Ayad et al. (1994) reported that enhanced adsorption of laminin on tissue-culture polystyrene promoted subsequent endothelial cell adhesion. These studies provided evidence that adsorption of specific protein(s) can, subsequently, control select cell adhesion on material surfaces. 

Steele, J. G, McFarland, C., Dalton, B. A., Johnson, G, Evans, M. D. M., Howlett, C. R., and Underwood, P. A., Attachment of human derived bone cells to tissue culture polystyrene and to unmodified polystyrene The effect of surface chemistry upon initial cell attachment. /. Biomat. Sci. Polymer Ed. 5, 245-257 (1993). 

We have used the coUagen-glycosaminoglycan (coUagen-GAG) mesh scaffold to probe molecular level cell-biomaterials interactions [11]. We seeded IMR-90 human fibroblasts onto three-dimensional (3-D) collagen-GAG meshes and control surfaces of tissue culture polystyrene (TCPS). Nucleic acids (mRNA) from cells from each culture were isolated, amplified, and hybridized to human genome microarrays (U133A Gene Chip, Affymetrix, Santa Clara, CA). 

Figwe 7 Proliferation data (days 1-3) and fluorescence microscopy images (day 1) of NIH-3T3 cells on control tissue culture polystyrene (TOPS) and on Aims formed from RLP12. The scale bar represents 100 pm. Reproduced with permission from Charati, M. B. Ifkovits, J. L. Burdick, J. A. etal. Soft MatterZOOS, 5, 3412. Copyright 2009 Royal Society of Chemistry. 

Looking to establish structure-property relationships in a different biological context, Kohn et al and Effah-Kaufmann and Kohn also examined the impact of their polyarylate library on the proliferation of UMR-106 osteoblast-like cells and fibrinogen adsorption as a measure of polymer biocompatibility. An analysis of total DNA concentration as well as metabolic and alkaline phosphatase activity after exposure of UMR-106 osteoblast-like cells to polyarylates revealed no noticeable cytotoxicity, with alkaline phosphatase and metabolic activities equivalent to that observed with tissue culture polystyrene. Polymers containing short ethyl ester side chains stimulated cell proliferation more effectively than more hydro-phobic ester side chains, while no correlations were observed between cell growth and strurtural features of the polymer backbone. 

Zhou et al. [14] and then Garner et al. [115,107] successfully polymerized PPy containing heparin as the dopant and evaluated its suitability for use as a substrate to support endothelial cell proliferation. When PPy was grown in the presence of heparin the resulting polymer can sustain the attachment and proliferation of human umbilical vein endothelial cells (HUVEC). The HUVECs spread and adopted morphology typical of that grown under standard in vitro conditions on tissue culture polystyrene. However, when the PPy was grown in the presence of nitrate ions (NO3") the seeded HUVECs did not attach. This result was at least in part attributed to the increased hydrophobic nature of the PPy-N03 compared to the PPy-heparin polymer. 

Electrical stimulation has been shown to enhance nerve cell regeneration [124,125], the mechanisms for this effect are, however, unclear. One hypothesis is that an electrical stimulus alters the local electrical fields of extracellular matrix molecules, changing protein adsorption [126]. As early as 1994, studies into the suitability of ICPs such as PPy as neuronal scaffolds provided positive proof that these electroactive stimulus response polymers indeed have a role to play. Shastri et al. [127] showed that neurite extension of PC 12 cells was more pronounced on PPy surfaces as compared to tissue culture polystyrene. The authors also showed that the application of an electrical stimulus to the cell culture on the PPy film significantly increased the expression of neurites in the cells compared to the controls. In addition, they demonstrated through tissue compatibility and transected sciatic nerve regeneration studies in rat models that the PPy films invoke little negative response and support nerve regeneration. 

Figure 12. Population density (A) and adhesion area (B) of osteoblast-like MG 63 cells on day 2 after seeding on tissue culture polystyrene dish (TCPS). carbon fibrereinforced carbon composites (CFRC) and CFRC coated with a fullerene layer (CFRC+full). C Growth curves of MG 63 cells on a terpolymer of polytetrafluoroethylene. poljcvinyldifluoride and polypropylene (Ter), terpolymer mixed with 4 wt. % of single-wall carbon nanohorns (SWNH) or 4 wt.% of high crystalline electric arc multi-wall nanotubes (MWNT-A). D Growth curves of MG 63 cells on TCPS. a nanostructured diamond layer (Nano) and a layer with hierarchically organized micro-and nanostructure (Micro-Nano). Mean S.E.M. from 4-12 measurements. ANOVA. Student-Newman-Keuls method. Statistical significance TCPS. CFRC. Ter p<0.05 compared to the values on tissue culture polystyrene, pure CFRC and pure terpolymer [23].

Fig. 30 Mesenchymal stem cell (MSC) proliferation on the surface of MDx-NCn gels and tissue culture polystyrene (TCPS) on days 1, 4, and 7, as demonstrated by ATP assay (n = 3). The data represent mean standard deviation. Reprinted from Kotobuki et al. [132], Copyright 2013, with permission of Wiley...

The described assay (ELISA) is a semiquantitative technique and is based on a comparison of the spectroscopic absorption values with those obtained with reference materials. Tissue culture polystyrene (TOPS) is used as a reference surface with a high protein-binding capacity. The detection of the binding capacity of antibodies is useful to establish the amount of adsorbed and immobilized fi-bronectin. When a high amount of adsorbed or immobilized fibronectin exists... 

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