Experimental Models for B-Cell Development B-Cell Culture/ Bone Marrow Stromal Cell Culture Bone marrow stromal cells provide support in the form of cytokines and binding proteins B-cell development can be examined through this model Fluorescence Activated Cell Sorting ● FACs permits the study of specific B-cell subpopulation separated based off the presence or absence of molecular markers ● After sorting these cells can be cultured and reanalyed for developmental order ● Pro-B Cells: B220+ CD43+ ● Pre-B Cells: B220+ CD43- IgM- (IgM expressed, but not on surface) ● Immature B-Cells: B220+ IgM+ IgD-● Mature B-Cells: B220+ IgM + IgD+ PCR Assay for Sequence of Ig Gene Rearrangement ● After having the cells sorted, a simple PCR assay using specific V, D, J primers can determine whether rearrangement of the heavy/light chain occurred ● PCR product will only form if rearrangement has occurred otherwise the primers are too far away ● The same can be done for genes like RAG1/2, which are implicated in a
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This note was uploaded on 05/06/2008 for the course MCB 150 taught by Professor Coscey during the Spring '08 term at Berkeley.