1 reply to this topic

[margyn87]

Hi all!

Currently, I am differentiating hESC to trophoblast lineage..I do time course experiment in treating them with various signaling molecules. Currently, the longest time I treat them is 4 days.. However, I did not see any morphological differences between treated culture and untreated culture..
I wonder whether the treatment time that I use is too short? What is the common or average time for treatment in any differentiation protocol of any type of stem cells?
I hope all of you don't mind to share with me.. Since it's kind of new field for me..

Best regards

[eri_b]

Hi, I work with hESCs in a different differentiation model, but my differentiation protocol takes 12 days to complete, using a variety of different factors that are applied in different stages. Having said that, morphological changes are obvious very quickly. I don't know anything about trophoblasts, but I would just double check that the reagents you are using are still active. Furthermore, how are you propagating/passaging your ES cells? With hESCs, i've found that my cells would not differentiate properly unless I passaged them manually (not enzymatically) by chopping up colonies into smaller pieces and transferring them to new feeder layers.

Good luck!