why my BSA contained medium cannot nourish my cells - (Feb/02/2007 )
thanks mito1, sorry for forcing you to guess. I know such require is cruel. By my different test with different controls, I more and more sure that protocol in published paper, I could not repeat, I could not say there are something wrong in their protocol, but at least my cell dosnt like that, even we use the exactly same cell line as them. I repeat that 2 hour starvation step sveral times, after that 2 hours, cells all already partly died. So from 10%FCS to 1%BSA by just 2 hour starvation; at least my cells dont like and cannot adapt so quick!
So I will order the serumfree media and systematicly construct my own protocol now.
thanks
I am in france; always have to wait one day to get your advice.
Good luck, lilylille!
I don't mind having trying to figure out protocols, but I just worry if it is someone else who will suffer if I am wrong. I have run into problems too, trying to follow published papers (the results are never quite the same as the what the authors get
). It may be something as simple as the lot number of the BSA you are using!Let us know what happens
, and don't be afraid to keep asking!mito 1
hello mito1
I already searched some information about the serumfree media, in fact there is no product surely will fit my cell type, nobody really ever test on my cell type, but anyway form Invitrogen and HyClone I got their proposed media type, AIMV ans HyQ CCM1. The reason for their proposal is those media has been shown fit large variety of cell types, but for my cell both said no guanrantee. Anyway I will do that test if nobody did that before.
Now what I want to ask is when I got the serum free media, my cell normally gorw in 10% FBS media, I should do the gradiently reduced FBS media to culture my cell and then switch to new serumfree media ?? Or I can directly switch to the serum free media, for normally there is said no nutrition deficient problem for serum free media, right?
look forward to the suggestion for you or other experinced guys.
thanks a lot
I already searched some information about the serumfree media, in fact there is no product surely will fit my cell type, nobody really ever test on my cell type, but anyway form Invitrogen and HyClone I got their proposed media type, AIMV ans HyQ CCM1. The reason for their proposal is those media has been shown fit large variety of cell types, but for my cell both said no guanrantee. Anyway I will do that test if nobody did that before.
Now what I want to ask is when I got the serum free media, my cell normally gorw in 10% FBS media, I should do the gradiently reduced FBS media to culture my cell and then switch to new serumfree media ?? Or I can directly switch to the serum free media, for normally there is said no nutrition deficient problem for serum free media, right?
look forward to the suggestion for you or other experinced guys.
thanks a lot
Definitely wean your cells off the FBS media before switching to the SFM: don't forget, the serum (FBS) has ingredients that may not be found in the SFM, or may be in different amounts. The cells need time to get used to their new food source.
mito 1
NOW I want to order the serum free media from Invitrogen, but the new question comes.
one containing Human BSA , price 36 euro 500 ml and the other contain bovine BSA , price 85 euro 500 ml. There are big difference of price between two, please tell me how I should judge and choose good enough for me and not necessarily expensive.
Urgent, please help. My cell is from mouse, but that is what we should think to judge? WHQT the difference between human BSA or Bovine BSA?
THANKS A LOT