Thermally Reversible Hydrogel Sheets for Adult Stem Cell Culture

A simple non-invasive method of cell retrieval using Methylcellulose (MC) coated Tissue Culture Poly-Styrone (TCPS) dishes was developed to allow the creation of single and multilayered adult stem cell sheet constructs while preserving the cell-cell and the cell–extracellular matrices. The optimal combination of MC-water-salt was found to be 12 to 14% of MC (mol. wt. of 15,000) in water with 0.5x PBS (∼150 mOsm). This solution exhibited a gel formation temperature of ∼32°C. The addition (evenly spread) of 200μl of 2mg/ml bovine collagen type -I (pH adjusted to 7.5) over the MC coated surface at 37°C, significantly improved the adult stem cell (ASC) adhesion and proliferation on the hydrogel system.