Flow analysis on microcasting with degassed polydimethylsiloxane micro-channels for cell patterning with cross-linked albumin

PLoS One. 2020 May 20;15(5):e0232518. doi: 10.1371/journal.pone.0232518. eCollection 2020.

Abstract

Patterned cell culturing is one of the most useful techniques for understanding the interaction between geometric conditions surrounding cells and their behaviors. The authors previously proposed a simple method for cell patterning with an agarose gel microstructure fabricated by microcasting with a degassed polydimethylsiloxane (PDMS) mold. Although the vacuum pressure produced from the degassed PDMS can drive a highly viscous agarose solution, the influence of solution viscosity on the casting process is unknown. This study investigated the influences of micro-channel dimensions or solution viscosity on the flow of the solution in a micro-channel of a PDMS mold by both experiments and numerical simulation. It was found experimentally that the degassed PDMS mold was able to drive a solution with a viscosity under 575 mPa·s. A simulation model was developed which can well estimate the flow rate in various dimensions of micro-channels. Cross-linked albumin has low viscosity (1 mPa·s) in aqueous solution and can undergo a one-way dehydration process from solution to solid that produces cellular repellency after dehydration. A microstructure of cross-linked albumin was fabricated on a cell culture dish by the microcasting method. After cells were seeded and cultivated on the cell culture dish with the microstructure for 7 days, the cellular pattern of mouse skeletal myoblast cell line C2C12 was observed. The microcasting with cross-linked albumin solution enables preparation of patterned cell culture systems more quickly in comparison with the previous agarose gel casting, which requires a gelation process before the dehydration process.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Albumins
  • Animals
  • Cell Culture Techniques / methods*
  • Dimethylpolysiloxanes
  • Mice
  • Microtechnology / methods
  • Myoblasts
  • Sepharose

Substances

  • Albumins
  • Dimethylpolysiloxanes
  • baysilon
  • Sepharose

Grants and funding

This research was supported by RIKEN-AIST"challenge research" project, RIKEN BDR organoid project, JSPS Grant-in-Aid for Scientific Research on Innovative Areas(19H05338), and Sasakawa Scientific Research Grant (20192031).The funders had no role in study design,data collection and analysis,decision to publish,or preparation of the manuscript.