If I don’t see organoid growth after 4 days, should I start over or give the culture more time? How long should I wait before starting over?

Answer

You will almost always see growth within 4 days of standard culture. Different mouse models will introduce some variability into this time frame but one should be able to recognize growing epithelial cysts after 1-2 days. Living cysts can be identified by a monolayered epithelium when observed under a brightfield microscope compared to single cells of dead crypts. If you are unsure if you have living cysts, I encourage you to contact our Tech Support team – they can identify living cells in your culture if images are sent. Contact details can be found here.