Archived Protocol

<HTML> <HEAD> <META NAME="GENERATOR" CONTENT="Adobe PageMill 3.0 Mac"> <TITLE>Choosing medium for tissue culture cells</TITLE> </HEAD> <BODY BGCOLOR="#ffffff" LINK="#000094"> <BLOCKQUOTE><TABLE WIDTH="450" BORDER="0" CELLSPACING="0" CELLPADDING="0"> <TR> <TD WIDTH="100%"> <H2><TABLE WIDTH="400" BORDER="0" CELLSPACING="0" CELLPADDING="0"> <TR> <TD WIDTH="100%"> <H3>Bart's Cookbook--<HR ALIGN=LEFT></H3> </TD> </TR> </TABLE>Tissue Culture Media<HR ALIGN=LEFT></H2> <P>We use two different kinds of media. Most cells are grown in DMEM. A few lymphoid cell lines are grown in RPMI. Cells grown in DMEM must be grown in a 10% CO2 atmosphere. As a result, most of our incubators are set for this. In contrast, cells grown in RPMI must be grown in a 5% CO2 atmosphere. We have one incubator set up for this. If you grow cells in RPMI in 10% CO2, the medium will be too acidic (yellow).</P> <H4>DMEM.</H4> <P>This is an abbreviation for Dulbecco/Vogt modified Eagle's (Harry Eagle) minimal essential medium. We use commercially-made DMEM as the medium for most cells. DMEM differs from the original MEM in that it contains approximately 4 times as much of the vitamins and amino acids present in the original formula, and some (2 to 4 fold) more glucose. It also contains iron and a few other oddments. These modifications were made by Marguerite so she did not have to come in on Sunday to change the medium on her cells.<BR> <BR> Most kinds of cells--human, monkey, hamster, rat, mouse, chicken--grow well in this medium.</P> <H4>RPMI.</H4> <P>This is an abbreviation for Roswell Park Memorial Institute medium. Human lymphoid cells are traditionally grown in RPMI medium. This is made from a mix by Margie. This medium contains a great deal of phosphate and is formulated for 5% CO2.<BR> <BR> Medium is always supplemented with serum.<BR> <BR> Most cell lines are grown in DMEM supplemented with either 10% calf serum or 10% fetal calf serum. Transformed cells usually grow well in calf serum. Normal 9non-transformed) cells sometimes require fetal calf serum, which is richer. Some clones of 3T3 cells do noticeably better in fetal calf serum. Some very fastidious cells are grown in 20% fetal calf serum. <BR> <BR> Due to the cost and scarcity of fetal calf serum, some cells are grown in horse serum instead.<BR> <BR> The medium of some lymphoid cells is supplemented with 5 x 10-5 M 2-mercaptoethanol. It does not matter whether this has undergone oxidation.<BR> <BR> Some cells benefit from the inclusion of 2 mM glutamine, 2, 5 or 10% tryptose phosphate broth, 2 mM pyruvate, or non-essential amino acids in their media. For example, Mary-Ann Campbell always puts pyruvate and non-essential amino acids, in addition to mercaptoethanol, in the medium of her B cells.<BR> <BR> <A HREF=http://pingu.salk.edu/~sefton/Hyper_protocols/TableOfContentsTC.html>Back to Table of Contents</A> </TD> </TR> </TABLE></BLOCKQUOTE> </BODY> </HTML>