I have issues with bad cell attachment when transferring my pluripotent stem cells from feeder cells or from feeder-free culture substrates to laminin. What can I do?
If the transfer from other culture substrates is problematic, please follow the instruction for transfer to Biolaminin. Some of the aspects are listed here:
- We recommend transferring the cells as single cells (or as small clumps) and always with the addition of apoptosis inhibitor, such as ROCKi for the first few (3-5) passages. Once the cells are adapted to the Biolaminin matrix, the cells can be cultured as single cells without ROCKi. This may take up to 5 passages – give it some time.
- If the cells are hard to adapt, try increasing the coating concentration to 10 µg/ml and seed at a higher cell density 50 000–100 000 cells/cm2. Once the cells are adapted a lower coating and seeding concentration often can be used.
- It is important that the cells transferred to the Biolaminin matrix are of high quality. Biolaminin 521 also supports specialized cell types so carefully select only undifferentiated cell areas for transfer.
- Biolaminin 521 works well in combination with most commercial media brands. However, it is to be expected that cell morphology will look different depending on the medium used for culture.