Micropatterned Smart Culture Surfaces via Multi‐Step Physical Coating of Functional Block Copolymers for Harvesting Cell Sheets with Controlled Sizes and Shapes

Abstract

Cell micropatterning on micropatterned thermoresponsive polymer‐based culture surfaces facilitates the creation of on‐demand and functional cell sheets. However, the fabrication of micropatterned surfaces generally includes complicated procedures with multi‐step chemical reactions. To overcome this issue, this study proposes a facile preparation of micropatterned thermoresponsive surfaces via a two‐step physical coating of two different diblock copolymers. Both copolymers contain poly(butyl methacrylate) blocks as hydrophobic anchors for water‐stable polymer deposition. At first, thermoresponsive polymer layers are constructed on cell culture dishes via spin‐coating block copolymers containing poly(N‐isopropylacrylamide) blocks that exhibit a transition temperature of ≈30 °C in aqueous media. To create polymer micropatterns on the thermoresponsive surfaces, microcontact printing of block copolymers containing hydrophilic poly(N‐acryloylmorpholine) (PNAM) blocks is performed using polydimethylsiloxane stamps. Stamped PNAM‐based block polymers are adsorbed to the outermost thermoresponsive surfaces, and increase the surface hydrophilicity with decreasing protein adsorption. Cells adhere and proliferate on the thermoresponsive domains at 37 °C, whereas the stamped hydrophilic domains remain cell‐repellent for 7 days. At 20 °C, cell sheets with controlled sizes and shapes are harvested from the surfaces with the desired micropatterns. This technique is useful for the preparation of micropatterned polymer surfaces for various biomedical applications.

Advances in the design, generation, and application of tissue-engineered myocardial equivalents

“…Thermoresponsive polymers, as poly(N-isopropylacrylamide), found use to generate sheets of CMs, CFs, SMCs, ECs, and so on. Differently from other biomaterials-based strategies, this approach has the unique advantage of generating a complete tissue by combining several cell sheets, including the ones needed to reconstruct a vascular net able to prevent inadequate perfusion and core necrosis ( Masuda et al, 2015 ; Iwamiya et al, 2016 ; Kawamura et al, 2017 ; Nagase et al, 2017 ; Nakayama et al, 2021a ; Nakayama et al, 2021b ). Intriguingly, thermoresponsive polymer technology has been demonstrated to be a useful tool to create biological pumps by shaping multilayered cell sheets in beating tubes or ventricles ( Mohammadi et al, 2022 ; Sekine and Okano, 2022 ).…”

Harvesting methods of umbilical cord-derived mesenchymal stem cells from culture modulate cell properties and functions

“…PNIPAAm exhibits reversible hydration/dehydration changes in response to cell culture temperatures near 37 °C [ 30 ]. The unique properties of PNIPAAm have been exploited in the context of biomedical applications such as drug delivery systems [ [31] , [32] , [33] , [34] , [35] , [36] , [37] ], bioanalysis and biosensor devices [ [38] , [39] , [40] , [41] , [42] , [43] , [44] ], nano-actuators [ [45] , [46] , [47] , [48] ], bioseparation tools [ [49] , [50] , [51] , [52] , [53] , [54] ], cell-separation materials [ [55] , [56] , [57] , [58] , [59] ], cell culture substrates [ [60] , [61] , [62] , [63] , [64] , [65] , [66] , [67] , [68] , [69] ], and cell sheet therapy in diverse types of regenerative medicine [ [16] , [17] , [18] , [19] , [20] , [21] , [22] , [24] , [25] , [26] , [27] , [28] ]. For cell sheet preparation, the cells were seeded onto PNIPAAm-modified dishes.…”