Steve V. John Morgridge Institute for Research-Metabolism, University of Wisconsin-Madison

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Jing Fan University of Wisconsin-Madison, 2017-
1 protocol

Gretchen Seim Nutritional Sciences, University of Wisconsin - Madison, 2017-
1 protocol

Steve V. John

Graduate Student, Morgridge Institute for Research-Metabolism, University of Wisconsin-Madison

Research focus

1 Author merit
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Education

BS, The College of New Jersey, USA, 2014

1 Protocol published

Metabolomic and Lipidomic Analysis of Bone Marrow Derived Macrophages

Authors: Gretchen L. Seim, Steven V. John and Jing Fan, date: 07/20/2020, view: 4796, Q&A: 1

Macrophages are highly plastic immune cells that are capable of adopting a wide array of functional phenotypes in response to environmental stimuli. The changes in macrophage function are often supported and regulated by changes in cellular ...

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Megan Phifer

Duke University

Hi all, I am wondering whether the density of the cells cultured in the 6-well plate is related to the size of the cell. In my case, the cells I am gonna use are smaller than macrophages. I am wondering whether I need to put in more cells per well in a 6-well plate during the preparation stage or still keep it at a density of 5*10^5 cells/well. What are your suggestions?

6/21/2022 8:39:20 AM Reply

Steve John

Morgridge Institute for Research-Metabolism, University of Wisconsin-Madison

Macrophages expand a lot in morphology when we stimulate them. 5e5 works for us because the macrophages still maintain a good confluency and we get good metabolite signals. If you are using a smaller cell type, a higher density definitely makes sense. But I would recommend trying a few different plating densities to determine the optimal seeding for your analysis. Hope this helps!

2022-06-23 15:27:20