Qustions about stem cell culture - (Feb/08/2006 )

Hi, Dear All,

I have a stupid question to show. I am prepareing culturing some stem cell. As I have no experience, I am a little nervse. I have two questions:

1) Usually we add serum to the medium directly without filter or any sterilize. But sombody tell me that when he culture stem cell, he will filter everything in the medium(including serum and other supplements). Is that reasonable because of the sensitivity of the stem cell.


2) Do the quality of MEF feeder cell will affect the growth of stem cell? Last time I treat the feeder cell with trypsin around 20 min for some reason, I am very worried about the cells condition. What is more, how many passages can the MEF feeder cell be continued? It seems the more passage, the cells grow more slow.

Thank you guys for any of nice suggesiton!

1)usually, we use commercial FBS and DMEM and other reagents from invitrogen,which is sterile and can be directly used.
2)arter digetion and pass the stem cell and MEF mixture onto new feeder-layer,the irradiated or Mitomycin treated MEF can't divide and then die,leaving stem cells alive on the new feeder.

Hi, Dear All,

I have a stupid question to show. I am prepareing culturing some stem cell. As I have no experience, I am a little nervse. I have two questions:

1) Usually we add serum to the medium directly without filter or any sterilize. But sombody tell me that when he culture stem cell, he will filter everything in the medium(including serum and other supplements). Is that reasonable because of the sensitivity of the stem cell.


2) Do the quality of MEF feeder cell will affect the growth of stem cell? Last time I treat the feeder cell with trypsin around 20 min for some reason, I am very worried about the cells condition. What is more, how many passages can the MEF feeder cell be continued? It seems the more passage, the cells grow more slow.

Thank you guys for any of nice suggesiton!