hTEC/SVTERT24-B Human thymic epithelial Cell Line (For-Profit use)

In vitro propagation

Passaging of cells

The new culture flasks have to be pre-coated with human collagen I. Therefore, the culture flasks are treated with Collagen I solution (80 µl/cm²) at 37°C for at least 30 min. Before introducing cells, remove excess of Collagen I solution and rinse flask once with PBS (160 µl/cm²).

For detachment of the cells, remove and discard the culture medium and wash the cells once with PBS. Remove PBS completely.

Then, add 0.05 % Trypsin-EDTA solution (RT, 20 µl/cm², Gibco, Cat# 25300-054), make sure that all cells have been in contact with this solution and incubate the culture flask at 37°C for approximately 1-3 min. Observe cell detachment under an inverted microscope. As soon as all cells are detached (if necessary agitate the cells by gently hitting the flask), add Defined Trypsin Inhibitor (20 µl/cm²; Gibco, Cat# R007100). Thereafter, resuspend the cells in growth medium and centrifuge at 170 g for 5 min. Discard the supernatant, resuspend the cell pellet in the remaining droplet and add growth medium (about 160 µl/cm²). Then, add appropriate aliquots of the cell suspension to new culture vessels supplemented with growth medium (final volume of 240 µl/cm²). A split ratio of 1:6 to 1:8 twice a week is recommended after having reached about 90 % confluence. Never allow the culture to become completely confluent! Cultivate cells at 37°C in a humidified atmosphere with 5% CO2.

Cryopreservation