Cell Capture and Culture Through Microtextured and Surface Treated Soft Polymers

The advancement of cell capture and culture technology is pivotal for progressing biological research and biomanufacturing. For example, the extraction and culture of primary cells from tissues - a procedure that has become increasingly useful with the advent of drug testing, regenerative medicine and personalized medicine. Tissue biopsies or surgical resections are the primary source of human cells for patient derived organoids (PDOs) and are gold-standards in cancer diagnosis, though they exhibit considerable pitfalls including: scarring and bleeding, discomfort and pain during and after the procedure, capture of epithelial cells which can overgrow desired cells in the PDOs.<br/><br/>Another area of interest concerns cell passaging: an often manual, time-intensive, and waste-producing process. In this process, cells are de-attached from the surface, usually using an enzymatic method like trypsin, diluted in media and plated onto new surfaces, either expanding or maintaining the cell culture. Trypsin and other enzymes like accutase are known to cause changes in the proteome of passaged cells, necessitate neutralization with inhibitors which adds steps and contamination risks, might affect the properties and the growth of primary cells on synthetic substrates during culture, and exhibit batch-to-batch variability affecting experimental consistency. The use of these enzymes also involves more steps, elapsed time and waste.<br/><br/>To combat these pitfalls, in this work, we develop a novel microtextured and surface-treated soft PDMS cell capture surface, designed to address these challenges by providing a more efficient, less invasive, and enzyme-free method of cell detachment and capture. By pressing the soft microstructured surface onto cell laden surfaces, cells are captured onto the surface. These can then be removed and re-cultured. Experimental results show an optimal pressure and surface design for maximizing viable cell capture. Through a combination of biomarkers, scanning electron microscopy (SEM) imaging, and in-situ live imaging, a surface chemistry mechanism of cell de-adhesion and capture is proposed. Finally, cells are successfully captured and cultured, showing the application of this technology.