Cell immobilization on patterned substrate
keywords: cellular growth, microcontact printing, ECM
Mastering the adhesion landscape of living cells on a surface is emerging as a new tool for investigating a lot of fundamental mechanisms of cell biology such as shape control, differentiation, division, polarity or motility. The common point of all these studies is the production of micro-patterns of various shapes and dimensions along which, adherent cells are immobilized in a deterministic way. The control of cell adhesion through technology allows the biologists to design specific scenarii and record how various living cells respond (neural, epithelial, tumoral and stem cells). Because technology allows reproducing identical precise patterns along well arranged periodical arrays, these experimental observations can be made systematically over large population of cells thus reaching a high level of representativeness. The micro-contact printing of proteins of the Extra Cellular Matrix (ECM) on conventional glass slides or coverslips turned out to be an efficient method for fabricating these micro-patterned cell culture substrates.
Biosoft has developed an instrument (Innostamp40) and a dedicated protocol enabling the automated production of highly reproducible functional micro-patterns in order to bring this process to a level where it could become a standard in cell biology. As seen in figure 1, adherent cells can be immobilized on-demand on the printed patterns.
Figure 1:fluorescence images of PC3-GFP cells immobilized on ECM micro patterns of various shapes (the patterns are depicted by dashed lines).
The workflow of this study is based on 3 steps (figure 2).
figure 2:the workflow of Cellular growth on patterned samples
The first one is the fabrication of ECM (Extra Cellular Matrix) protein micro patterns on glass slide. For that, we used the automatic microcontact printer named InnoStamp40. It is based on magnetic field assisted microcontact printing. This technology allows:
- An automation of the full process of microcontact printing,
- A fine control of the pressure applied during the printing step,
- An alignment of the patterns on the sample,
- A high reproducibility of the process.
Three elements are manipulated into the InnoStamp40® (www.innopsys.com):
- A PDMS stamp with the micropatterns,
- A sample (in this study: a glass slide),
- Some ink (In this study, it is a 100µg/mL ECM protein solution with PBS 1X).