Control of adhesion of human induced pluripotent stem cells to plasma-patterned polydimethylsiloxane coated with vitronectin and γ-globulin

Abstract

Human induced pluripotent stem cells (hiPSCs) are a promising source of cells for medical applications. Recently, the development of polydimethylsiloxane (PDMS) microdevices to control the microenvironment of hiPSCs has been extensively studied. PDMS surfaces are often treated with low-pressure air plasma to facilitate protein adsorption and cell adhesion. However, undefined molecules present in the serum and extracellular matrix used to culture cells complicate the study of cell adhesion. Here, we studied the effects of vitronectin and γ-globulin on hiPSC adhesion to plasma-treated and untreated PDMS surfaces under defined culture conditions. We chose these proteins because they have opposite properties: vitronectin mediates hiPSC attachment to hydrophilic siliceous surfaces, whereas γ-globulin is adsorbed by hydrophobic surfaces and does not mediate cell adhesion. Immunostaining showed that, when applied separately, vitronectin and γ-globulin were adsorbed by both plasma-treated and untreated PDMS surfaces. In contrast, when PDMS surfaces were exposed to a mixture of the two proteins, vitronectin was preferentially adsorbed onto plasma-treated surfaces, whereas γ-globulin was adsorbed onto untreated surfaces. Human iPSCs adhered to the vitronectin-rich plasma-treated surfaces but not to the γ-globulin-rich untreated surfaces. On the basis of these results, we used perforated masks to prepare plasma-patterned PDMS substrates, which were then used to pattern hiPSCs. The patterned hiPSCs expressed undifferentiated-cell markers and did not escape from the patterned area for at least 7 days. The patterned PDMS could be stored for up to 6 days before hiPSCs were plated. We believe that our results will be useful for the development of hiPSC microdevices.

Hydroxyapatite Nanoparticle Coating on Polymer for Constructing Effective Biointeractive Interfaces

“…Figure 5(b) is followed by the functionalization of the surface with γ-globulin and vitronectin and the interactions with human-induced pluripotent cells (hiPSCs). It was found that γ-globulin on the untreated PDMS surfaces blocked the vitronectin adsorption, which prevents hiPSC adhesion [177]. It was reported that allylamine plasma deposition on poly(D,L-lactic acid) (PDLLA) promoted N3T3 fibroblast adhesion and proliferation.…”

Modulation of human mesenchymal and pluripotent stem cell behavior using biophysical and biochemical cues: A review

“…More recently, new reagents such as small molecules which target specific intracellular pathways have been proposed for stem cell differentiation protocols, and are now widely used in a soluble form in culture media (Xu et al, ). At the same time, studies have focused on immobilization or pre‐coating bioactive signals on cell culture substrates or the surface of implantable scaffolds (Bacakova et al, ; Chen et al, ; Chuah et al, ; Duffy et al, ; Kang et al, ; Kuddannaya et al, ; Lu et al, ; Nagaoka et al, ; Yamada et al, ). This has the potential to establish more defined culture systems and be used in future clinical applications, such as commercialized cell culture tools (Higuchi et al, ; Wang et al, ) and tissue engineered implants (Salerno et al, ).…”

“…Direct surface modification is another method that has been used to influence cell behavior. Protein coatings, such as Type I collagen (Chuah et al, 2015;Kuddannaya et al, 2013) or fibronectin (Kuddannaya et al, 2013;Wang et al, 2012e;Yamada et al, 2014), can be applied to facilitate attachment of MSCs and iPSCs to hydrophobic surfaces like polydimethylsiloxane (PDMS). Different surface properties provided by these modifications can affect the density and configuration of protein adsorption prior to cell culture or from media, which in turn affects cell adhesion and growth (De Luca et al, 2015).…”

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

“…The patterning of cells, including hiPSCs, can be easily performed by masked plasma oxidation, which uses patterned physical masks to partially prevent plasma treatment. [7][8][9][10] Using masked plasma oxidation, we succeeded in creating hiPSC patterns (2-mm-diameter discs) on polydimethylsiloxane (PDMS) surfaces under culture conditions defined in a previous study. 9 PDMS is one of the most popular biocompatible materials for research and development of small cell culture devices because this elastomer is nontoxic, chemically inert, transparent, and gas permeable.…”

“…[7][8][9][10] Using masked plasma oxidation, we succeeded in creating hiPSC patterns (2-mm-diameter discs) on polydimethylsiloxane (PDMS) surfaces under culture conditions defined in a previous study. 9 PDMS is one of the most popular biocompatible materials for research and development of small cell culture devices because this elastomer is nontoxic, chemically inert, transparent, and gas permeable. [11][12][13] Plasma treatment on PDMS oxidized ≡Si-CH 3 to generate ≡Si-O-Si≡ groups suggests that hydrophilic and siliceous layers were formed on the surface 9,11 (Fig.…”