Cryopreservation of anchorage-dependent mammalian cells fixed to structured glass and silicon substrates

This paper describes a procedure for the cryopreservation of anchorage-dependent cells in a predefined position on microstructured glass or silicon substrates. During freezing and thawing, cells retain their location on the substrate, and an individual comparison and identification of cells before and after preservation are possible. To utilize this advantage, a good adherence and a high survival rate are important. It can be shown that adhesion of mouse fibroblasts (NIH-3T3) to substrate strongly influences the survival rate: 94% of cells grown for 16 h before freezing were judged to be alive after thawing. Widely spaced cells are best suited to cryopreservation on substrates. The different patterns of adhesion of cells to substrates when incubated for 1, 3, 6, and 16 h, were visualized by total internal reflection microscopy (TIRM).