The purification of B-cell precursors from mouse fetal liver.
Lymphocyte progenitor cells isolated on sequential days of gestation from mouse fetal liver represent distinct stages in B cell development. We have utilized polymerase chain reaction (PCR)-based assays to detect immunoglobulin (Ig) gene rearrangement and flow cytometry to assay cell surface markers following fractionation based on the differential expression of the B cell-specific phosphatase CD45 (B220). The purification of B220+ cells from day 17 fetal liver resulted in a 10-fold enrichment of cells which had undergone gene rearrangement events. We have also shown that day 13 fetal liver cells activate successive Ig gene rearrangements during short-term culture in the presence of fetal calf serum (FCS), interleukin-3 (IL3), and interleukin-7 (IL7). However, partially purified lymphocyte precursors fail to activate Ig gene rearrangement in culture unless they are cultured in the presence of a stromal cell line.
Morrow
T
T
Department of Medicine, John Hopkins University School of Medicine, Baltimore, Maryland 21205.
Schlissel
M
M
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology; D002469:Cell Separation; D002478:Cells, Cultured; D005333:Fetus / Q000166:cytology; D015322:Gene Rearrangement, B-Lymphocyte; D006412:Hematopoietic Stem Cells / Q000166:cytology; D008099:Liver / Q000166:cytology; D051379:Mice; D008807:Mice, Inbred BALB C