Plasma-Patterned Polydimethylsiloxane Surface With Single-Step Coating with a Mixture of Vitronectin and Albumin Enables the Formation of Small Discs and Spheroids of Human Induced Pluripotent Stem Cells

Abstract

Plasma treatment is an easy method of cleaning and hydrophilizing glass and polymer surfaces to facilitate adsorption of cell-adhesive proteins. Thus a patterned plasma treatment is useful for fabricating cell adhesion patterns. We previously succeeded in producing human induced pluripotent stem cell (hiPSC) discs (2 mm in diameter) on plasma-patterned polydimethylsiloxane (PDMS) by single-step coating of a mixture of 2 ubiquitous proteins, vitronectin and γ-globulin. Under serum-free and feeder-free conditions, without any undefined cell adhesion molecules, γ-globulin blocked hiPSC adhesion on surfaces not treated with plasma. However, γ-globulin has low cost-effectiveness and availability, and the resulting disc diameter was too large for cell-based assays. We demonstrate that bovine serum albumin (BSA) can also be used to block hiPSC adhesion on plasma-untreated PDMS surfaces coated with vitronectin. We succeeded in creating small hiPSC discs (200 μm in diameter) using single-step coating of a mixture of vitronectin and BSA. The hiPSCs proliferated without escaping from the patterned area and finally spontaneously detached from the discs to form spheroids. We believe that our method for generating hiPSC discs and spheroids will be useful for developing new bioengineering devices to enhance cell differentiation and to test drug safety for human embryonic development, contributing to future medical applications.

Application of Microfluidics in Stem Cell Culture

“…The patterned PDMS could be stored for up to 6 days before hiPSCs were plated. Furthermore, we demonstrate that not only γ-globulin but also bovine serum albumin (BSA) could be used to block human iPS cell adhesion on plasma-untreated PDMS surfaces ( Figure 2) [48]. The hiPSCs proliferated without escaping from the patterned area and finally detached spontaneously from the discs to form spheroids.…”

“…We succeeded in forming human iPS cells pattern on the PDMS surface by a simple technique using plasma 2 oxidation with perforated mask and defined culture conditions [48,49]. As described above, PDMS is one of the most popular biocompatible materials for research and development of cell culture microdevices.…”

Strategies for surface coatings of implantable cardiac medical devices

“…In this method, self-assembled albumin is designed to avoid unfordable or weak chemistry between the polymer chain and the albumin, using an intermediate which is a linker, the linker will provide a strong attachment of the albumin, but this technique still needs to be optimized to be applied in CMDs commercially (see Table 2 ) ( Gonçalves et al, 2011 ; Tao et al, 2019 ; Kuten Pella et al, 2022 ). Albumin coatings are mainly designed for polymeric materials of CMDs (e.g., polytetrafluoroethylene, polydimethylsiloxanes, polyurethane, polycarbonate), but at the moment, these coatings need to be improved to be applied in medical devices or CMDs ( Yamada et al, 2014 ; Cheng et al, 2019 ). One approach to solve this inconvenience is the combination of multiple compounds (e.g., heparin, elastin, and fibrinogen) linked with albumin to produce a suitable matrix ( Manabe and Nara, 2021 ).…”