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The need for basement membrane extract (BME) with undefined constituents, such as Matrigel, for intestinal stem cell (ISC) culture in traditional systems poses a significant barrier that must be overcome for the development of clinical-grade, scalable, ready-to-use ISCs. Here, we propose a functional polymer-based xenogeneic-free dish for the culture of intestinal stem cells (XF-DISC), ensuring substantially prolonged maintenance of ISCs derived from 3-dimensional human intestinal organoids (ISCs 3D-hIO ). XF-DISC enables remarkable expandability, exhibiting a 24-fold increase in cell numbers within 30 days, with long-term maintenance of ISCs 3D-hIO for more than 30 consecutive passages (>210 days). In addition, XF-DISC is fully compatible with a cell banking system. Notably, human pluripotent stem cell-derived ISCs 3D-hIO cultured on XF-DISC are successfully transplanted into intestinal injury and inflammation mouse models, leading to engraftment and regeneration of damaged mouse intestinal epithelium. As a reliable and scalable xenogeneic-free ISC 3D-hIO culture method, XF-DISC is highly promising for the development of regenerative ISC therapy for human intestinal diseases., Competing Interests: Competing interests: The authors declare no competing interests. Ethical considerations of working with human cells and animals: This study complies with all relevant ethical regulations. All studies based on human pluripotent stem cells were approved by the Korean Public IRB (IRB number: P01-201409-ES-01-09, P01-201609-31-002). Male NIG mice (Nod/scid-IL2Rgem1/GH, 6–12 weeks old; GHBio, Daejeon, Korea) were housed in a specific pathogen-free facility with a controlled 12-hour light/dark cycle, temperature maintained at 20-22 °C, and relative humidity at 55 ± 5%. The mice were provided with ad libitum access to water and fed a purified AIN-76A diet (Teklad Custom Research Diet, CA.170481; Envigo, Indianapolis, IN, USA). All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of KRIBB (KRIBB-AEC-22233). For the study based on human blood, informed consent was obtained prior to participation with approval from the IRB at Yonsei University Health System (IRB No. 3-2023-0347)., (© 2024. The Author(s).)

The shear-rolling process is a promising directed self-assembly method that can produce high-quality sub-10 nm block copolymer line-space patterns cost-effectively and straightforwardly over a large area. This study presents a high temperature (280 °C) and rapid (~0.1 s) shear-rolling process that can achieve a high degree of orientation in a single process while effectively preventing film delamination, that can be applied to large-area continuous processes. By minimizing adhesion, normal forces, and ultimate shear strain of the polydimethylsiloxane pad, shearing was successfully performed without peeling up to 280 °C at which the chain mobility significantly increases. This method can be utilized for various high-χ block copolymers and surface neutralization processes. It enables the creation of block copolymer patterns with a half-pitch as small as 8 nm in a unidirectional way. Moreover, the 0.1-second rapid shear-rolling was successfully performed on long, 3-inch width polyimide flexible films to validate its potential for the roll-to-roll process., (© 2023. The Author(s).)

Sub-10 nm patterns prepared by directed self-assembly (DSA) of block copolymer (BCP) thin films offer a breakthrough method to overcome the limitations of photolithography. Perpendicular orientation of the BCP nanostructures is essential for lithographic applications, but dissimilar surface/interfacial energies of two blocks generally favour parallel orientations, so that the perpendicular orientation could only be obtained under very limited conditions. Here, we introduce a generalized method for creating perpendicular orientations by filtered plasma treatment of the BCP films. By cross-linking the surface of disordered BCP films using only physical collisions of neutral species without ion bombardment or UV irradiation, neutral layers consistent with the BCP volume fraction are produced that promote the perpendicular orientations. This method works with BCPs of various types, volume fractions, and molecular weights individually at the top and bottom interfaces, so it was applied to orientation-controlled 3D multilayer structures and DSA processes for sub-10 nm line-spacing patterns.