cell culture protocol (URGENT) - (Aug/18/2008 )
Dear all
I will start to work with sarcoma cell lines (Saos-2 cell), anybody has experience with this cell line? I will be using DMEM with 10% FBS, do I need to add other reagents such as penstrep, fungizone, uridine, sodium pyruvate? If yes, can anybody help me how many Ul (micro) should I use?
I only had had experience with human fibroblast cell lines many years ago, couldn't remember it in detail.
And how to count the cells using haemocytometer?
And the other thing is, for the freezing medium, is it 90% FBS and 10% DMSO?
Thanx a lot
-neny-
QUOTE (neny @ Aug 18 2008, 09:26 PM)
Dear all
I will start to work with sarcoma cell lines (Saos-2 cell), anybody has experience with this cell line? I will be using DMEM with 10% FBS, do I need to add other reagents such as penstrep, fungizone, uridine, sodium pyruvate? If yes, can anybody help me how many Ul (micro) should I use?
I only had had experience with human fibroblast cell lines many years ago, couldn't remember it in detail.
And how to count the cells using haemocytometer?
And the other thing is, for the freezing medium, is it 90% FBS and 10% DMSO?
Thanx a lot
I will start to work with sarcoma cell lines (Saos-2 cell), anybody has experience with this cell line? I will be using DMEM with 10% FBS, do I need to add other reagents such as penstrep, fungizone, uridine, sodium pyruvate? If yes, can anybody help me how many Ul (micro) should I use?
I only had had experience with human fibroblast cell lines many years ago, couldn't remember it in detail.
And how to count the cells using haemocytometer?
And the other thing is, for the freezing medium, is it 90% FBS and 10% DMSO?
Thanx a lot
900 ul of serum and 100 ul DMSO to freeze.
use 10% serum, 89% DMEM, 1% Penicilin/Streptpmycin
also see here:
http://catalog2.corning.com/Lifesciences/e...hInfoIndex.aspx
in order to count the cells there is a file you can download from this website, but I don't remember where exactly, try to find it here.
also, try to wash your cells everyday with 1XPBS until you become familiar with cell culture and how to handle cells. because people usually experience contamination in cell culture especially during the first days of learning.
-Curtis-