Establishment of human embryonic stem cell - (Mar/12/2006 )
Hi!
I am trying my best to establish a new human embryonic stem cell line.But now I have some questions about my experiment:
the immunosurgery was well be done,and the morphologic features of ICM looks very well. but They were failure to attach to the feeder. the media used was DMEM/F12+SR. Now i was not sure about the DMEM/F12+FBS+LIF is necessary for the establishment of human embryonic stem cell? or the activity of the ICM isolated had been lost?
thank you in advance! 
LIF give no contribution to the hES cells!!!
I am trying my best to establish a new human embryonic stem cell line.But now I have some questions about my experiment:
the immunosurgery was well be done,and the morphologic features of ICM looks very well. but They were failure to attach to the feeder. the media used was DMEM/F12+SR. Now i was not sure about the DMEM/F12+FBS+LIF is necessary for the establishment of human embryonic stem cell? or the activity of the ICM isolated had been lost?
thank you in advance!
[quote name='JerryLee' date='May 26 2006, 09:09 AM' post='53210']
LIF give no contribution to the hES cells!!!
But many articles reproted that LIF was added when FBS was an source of protein especially during the early propagation!
Human ES is different from mouse ES in that they are LIF-independent (LIF alone can not do the trick).....you may need to combine with other factors. So far, only the Wisconsin group (company)has successfully developed a serum-free defined medium for maintaining human ES pluripotency.