First I want to confirm the presence of plasma cell in bone marrow and spleen. Then I would like to collect those plasma cell. So any idea how I can proceed ?
Thank you
Dear colleague, It is known that plasma cells are present in bone marrow and spleen of mammals. You could use of techniques showing the presence of specific antibody producing cells (see attach) or other specific fluorescence techniques for showing high immunoglobulin concentrations in the cytoplasm of plasma cells. Good luck with your work, Willem
Hi Selma,
I would suggest to use flow cytometry, also because after you would like to collect them, that you could do by cell sorting. Here, I leave you the link to a paper in which we identified and sorted a subset of plasma cells. It is an extensive study in which you can find several ways to study plasma cells.
Good luck.
Andreia
Article LAG-3 Inhibitory Receptor Expression Identifies Immunosuppre...
Hi Selma,
In naive mouse, at steady state, you can identify plasma cells in both bone marrow and spleen by flow cytometry by CD138hiB220- (mature PC) and CD138hiB220+(immature PC). If you are doing immunization such as NP-KLH, and to look at NP-specific plasma cells you could further fix the cells and do intracellular staining with fluorescent-NP and IgG in order to see NP-specific IgG-secreting plasma cells. Also, you could do ELISPOT by coating NP-antigen (such as NP-BSA) and then add total spleen or bone marrow cells after red blood cell lysis, culture in 37 degree incubator overnight and the next add secondary anti-IgG or anti-IgM antibody to identify the numbers of NP-specific IgG or IgM secreting plasma cells.
To isolate plasma cells, if you are doing end-point study with plasma cells you can use miltenyi MACS beads anti-CD138 positive isolation. If you are doing transfer with those plasma cells you can sort them by CD138hiB220- and/or CD138hiB220+. If you have Blimp1-reporter (e.g. Blimp1-YFP) mouse, then you can sort YFP from both spleen and bone marrow using CD3-YFP+ to get rid of T cells because they are also labeled in these mice.
Hope this help.
Zhixin
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