A Thermoresponsive Coating for Non-Enzymatic Release of Strongly Adherent Cells from Flasks and Other Closed-Vessel Formats

Traditional cell culture techniques rely on enzymatic digestion for passaging, which introduces cost, cell stress pathway activation, potential contamination, and a reduction in cellular viability. With the damage perpetuated via repeated enzymatic-based harvests of cell lines, the hurdle for regulatory approval of stem cell and regenerative therapies increases. Thermoresponsive polymer coatings can reduce or eliminate the need for enzymatic digestion and improve cell line health. Current commercial options include coated petri dishes and multi-well plates, but no culture flask is yet widely available. Additionally, most of these products do not offer digestion-free passaging for strongly adherent cell lines such as mesenchymal stem cells (MSC). ISurTec has developed a thermoresponsive coating for T25 flasks that allows non-digestive cell release, optimized for strongly adherent cell lines. This coating will be expanded to other cultureware including larger and multi-layer flasks, and roller bottles, as well as dishes and multi-well plates. The thermoresponsive polymer is hydrophobic above 32°C and becomes hydrophilic at lower temperatures, spontaneously releasing the cells within 30 minutes when brought to room temperature. In comparison to standard culture flasks, our coating showed normal proliferation rates for multiple cell lines. Our coating releases 90% of human MSC (cord blood-derived) within 30 minutes as compared to 50% released on a competitor’s coating. Our coating also releases cells grown to 100% confluence instead of the 60-70% recommended limit of a competitor. ISurTherm can be applied to multiple cultureware substrates and is one of the only thermoresponsive culture flasks now available on the market.