Regulation of the Contribution of Integrin to Cell Attachment on Poly(2-Methoxyethyl Acrylate) (PMEA) Analogous Polymers for Attachment-Based Cell Enrichment

Abstract

Cell enrichment is currently in high demand in medical engineering. We have reported that non-blood cells can attach to a blood-compatible poly(2-methoxyethyl acrylate) (PMEA) substrate through integrin-dependent and integrin-independent mechanisms because the PMEA substrate suppresses protein adsorption. Therefore, we assumed that PMEA analogous polymers can change the contribution of integrin to cell attachment through the regulation of protein adsorption. In the present study, we investigated protein adsorption, cell attachment profiles, and attachment mechanisms on PMEA analogous polymer substrates. Additionally, we demonstrated the possibility of attachment-based cell enrichment on PMEA analogous polymer substrates. HT-1080 and MDA-MB-231 cells started to attach to poly(butyl acrylate) (PBA) and poly(tetrahydrofurfuryl acrylate) (PTHFA), on which proteins could adsorb well, within 1 h. HepG2 cells started to attach after 1 h. HT-1080, MDA-MB-231, and HepG2 cells started to attach within 30 min to PMEA, poly(2-(2-methoxyethoxy) ethyl acrylate-co-butyl acrylate) (30:70 mol%, PMe2A) and poly(2-(2-methoxyethoxy) ethoxy ethyl acrylate-co-butyl acrylate) (30:70 mol%, PMe3A), which suppress protein adsorption. Moreover, the ratio of attached cells from a cell mixture can be changed on PMEA analogous polymers. These findings suggested that PMEA analogous polymers can be used for attachment-based cell enrichment.

Blood-Compatible Polymer for Hepatocyte Culture with High Hepatocyte-Specific Functions toward Bioartificial Liver Development

“…Recently, we reported that intermediate water content influenced the amount of protein adsorption. 29 We reported that increasing intermediate water content decreased protein adsorption amounts. As compared to intermediate water content in hydrated PMEA, the contents in hydrated PTHFA are lower.…”

Design of Polymeric Biomaterials: The “Intermediate Water Concept”

“…We found that PMEA and its analogs can suppress the protein adsorption on the substrates according to their intermediate water contents (Figure 9). 106,107 Thus, integrin-dependent cell adhesion was weakened on PMEA analogs which possess comparatively high intermedi- ate water contents (e.g., PMEA, poly{2-(2-methoxyethoxy) ethyl acrylate} (PMe2A), and poly[2-{2-(2-methoxyethoxy)-ethoxy}ethyl acrylate] (PMe3A) with the increase of their intermediate water contents. 107,108 Integrins play an important role in cell adhesion.…”

“…106,107 Thus, integrin-dependent cell adhesion was weakened on PMEA analogs which possess comparatively high intermedi- ate water contents (e.g., PMEA, poly{2-(2-methoxyethoxy) ethyl acrylate} (PMe2A), and poly[2-{2-(2-methoxyethoxy)-ethoxy}ethyl acrylate] (PMe3A) with the increase of their intermediate water contents. 107,108 Integrins play an important role in cell adhesion. Thus, cell adhesion is expected to be suppressed when integrin-dependent cell adhesion is weakened via the suppression of protein adsorption on the substrates.…”

Integrin-independent Cell Adhesion Substrates: Possibility of Applications for Mechanobiology Research

“…68,70,71 Recently, we reported that substrates coated with poly(2-methoxyethyl acrylate) (PMEA) and its analogous polymers, which suppress protein adsorption, allowed the cells to adhere. [71][72][73][74] On PMEA-analogous polymer-coated substrates, the cells can adhere even in the presence of ethylenediaminetetraacetic acid (EDTA), an inhibitor of integrindependent cell adhesion. The few focal adhesions formed in cells cultured on PMEA-analogous polymer-coated substrates also indicate that the cells adhered via integrin-independent mechanisms.…”

“…The few focal adhesions formed in cells cultured on PMEA-analogous polymer-coated substrates also indicate that the cells adhered via integrin-independent mechanisms. 72,73 The cells exhibited different functions on PMEA-analogous polymers-coated substrates compared with conventional culture substrates (e.g., tissue culture polystyrene). In particular, HepG2 cells were able to adhere on a PMEAcoated substrate via integrin-independent mechanisms and form a round shape that increased hepatocyte-specific functions such as albumin expression.…”