Polydopamine-mediated Surface Modification Promotes the Adhesion and Proliferation of Human Induced Pluripotent Stem Cells

Abstract

With their abilities of self-renewal and pluripotency to differentiate into all three germ layers, human induced pluripotent stem cells (hiPSCs) are a promising cell source for cell-based drug and implant testing. However, the large-scale expansion and maintenance of hiPSCs requires following strict protocols. There is high demand for advanced cell culture systems capable of generating high-quality hiPSCs to meet application requirements. In this study, we probe the possibility of modifying polymeric substrates for maintaining the self-renewal and pluripotency of hiPSCs. Here, polydopamine (PDA) was employed to immobilize the Laminin 521 (LN521) onto the surface of polyethylene terephthalate (PET). An aqueous solution of dopamine with concentrations ranging from 0 to 2.0 mg/mL was applied on PET surfaces. These PDA-modified surfaces were further functionalized with LN521. Surface wettability was evaluated by measuring the water contact angle (WCA) and surface properties of the modified substrate were analyzed using an atomic force microscope (AFM). Initial hiPSC attachment (1h after seeding) and cell proliferation were evaluated by counting the total cell number. The maintenance of pluripotency was evaluated at designed time points. WCA of the PDA-LN521 surfaces gradually decreased from 62.1°±6.3° to 8.1°±2.9°. The maximum peak-to-valley height roughness (Rt) of those surfaces determined by AFM increased in a dopamine-concentration-dependent manner, ranging from 43.9±1.6 nm to 126.7±7.6 nm. The Young’s modulus of these surfaces was substantially increased from 0.98±0.36 GPa to 4.81±2.41 GPa. There was a significant enhancement (13.0±7.2% and 24.2±8.1%) of hiPSC adhesion on PDA-LN521 (dopamine concentration at 0.125 and 0.25 mg/mL). When increasing the dopamine concentration to 0.5 and 1.0 mg/mL, there was no further increase in hiPSC adhesion on PDA-LN521 surfaces. Moreover, hiPSC proliferation was remarkably enhanced on PDA-LN521 surface (dopamine solution at concentration from 0.125 to 1.0 mg/mL). Pluripotency of hiPSCs was not affected by PDA treatment. In conclusion, PDA-mediated surface modification is an effective approach for the robust expansion and maintenance of hiPSCs on polymer substrates.