Can EBs be formed without growing them in adherent culture?
I grew my EBs in suspension culture for 6 days and then as I saw their morphology under microscope, discarded them to extract their RNA and RT-PCR to check the expression of three germ layers genes.
EBs are perfectly grown in suspension cultures. Normally around one week of culture (EBs from mESC), they show a double layer (visceral endoderm outside and primitive ectoderm inside), with an empty space in the center that corresponds to apoptotic cells. We were able to maintain them in suspension for up to 5 weeks. For more details, see "Expression of regulatory genes for pancreas development during murine embryonic stem cell differentiation" by Mfopou et al. (2005) in the Int J Dev Biol. 2005;49(8):915-22.
Hi Nasibe,
I am agree with Josue. We can easily growthe EBin suspension culture nicely.
- Badges
- Science topic