Abstract
Three-dimensional (3D) culture bridges and minimizes the gap between in vitro and in vivo states of cells and various 3D culture systems have been developed according to different approaches. However, most of these approaches are either complicated to operate, or costive to scale up. Therefore, a simple method for stem cell spheroid formation and preservation was proposed using poly(D,L-lactic acid) porous thin film (porous nanosheet), which were fabricated by a roll-to-roll gravure coating method combining a solvent etching process. The obtained porous nanosheet was less than 200 nm in thickness and had an average pore area of 6.6 μm2 with a porosity of 0.887. It offered a semi-adhesive surface for stem cells to form spheroids and maintained the average spheroid diameter below 100 μm for 5 days. In comparison to the spheroids formed in suspension culture, the porous nanosheets improved cell viability and cell division rate, suggesting the better feasibility to be applied as 3D culture scaffolds.
Highlights
Two-dimensional (2D) cell culture on tissue plates is commonly performed in in vitro cell biology studies, allowing researchers to study how cells expand, behave under various stresses, proliferate and/or differentiate in response to stimuli
We reported for the first time that porous nanosheets prepared by a solvent etching method could support the adipose-tissue derived stem cell (ASC) spheroids for high-magnification imaging owing to its transparency (Suematsu et al, 2020)
Porous nanosheets were integrated from two immiscible polymers, PS and PDLLA (1:1 in weight) via a gravurecoating method (Figure 1A), followed by a solvent etching process with cyclohexane to remove the phase-separated PS regions (Figure 1B)
Summary
Two-dimensional (2D) cell culture on tissue plates is commonly performed in in vitro cell biology studies, allowing researchers to study how cells expand, behave under various stresses, proliferate and/or differentiate in response to stimuli. Nanosheet for 3D Spheroid Culture cell viability, cell secretomes and differentiation ability (Park et al, 2017). Embryonic stem cell spheroids have shown multiple functional capabilities in development and differentiation (McKee and Chaudhry, 2017). Mesenchymal stem cells cultured in 3D systems exhibit secretion of antiinflammatory factors (Bartosh et al, 2010), enhanced cell survival (Emmert et al, 2013) and the osteogenic differentiation ability for bone regeneration application (Yamaguchi et al, 2014)
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