In vivo maturation of pre-B cells derived from long-term cultured bone marrow.
Long-term culture of a nonadherent pre-B cell line was established from BALB/c bone marrow by the method of Whitlock and Witte. Adherent filler cells, also derived from bone marrow, were required for the growth of such pre-B cells. The surface phenotypes of the nonadherent cells determined by FACS analysis were negative for mu-chain, kappa-chain, and Ia, but 50% of nonadherent cells expressed a marker recognized by the monoclonal 14.8. Analysis of cytoplasmic immunofluorescent staining revealed the presence of cytoplasmic mu-chain but absence of kappa-chain. These pre-B cells did not respond in vitro to such mitogens as LPS or DxSO4 and were not stimulated by allospecific helper T cells. However, when such BALB/c derived pre-B cells were transferred to irradiated BDF1 mice, recovered spleen cells expressed mu-chain as well as the marker recognized by 14.8. Such in vivo passaged pre-B cells were able to respond to LPS in vitro. Moreover, the response observed was diminished markedly by anti-H-2d antiserum and complement (C) but not anti-H-2b and C, indicating that the responder cells were of donor rather than host origin. Because the reconstitution of irradiated mice was limited to cells of the B lymphocyte lineage, it would appear that cells recovered from in vivo passage are the progeny of in vitro cultured pre-B cells and not that of in vitro contaminating stem cells.
Nagasawa
R
R
Kanagawa
O
O
Tittle
T V
TV
Chiller
J M
JM
D000818:Animals; D001402:B-Lymphocytes / Q000145:classification / Q000166:cytology* / Q000276:immunology; D001854:Bone Marrow Cells; D016026:Bone Marrow Transplantation; D002454:Cell Differentiation; D002478:Cells, Cultured; D016264:Dextran Sulfate; D003911:Dextrans / Q000494:pharmacology; D005260:Female; D007116:Immunization, Passive; D008070:Lipopolysaccharides / Q000494:pharmacology; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D010641:Phenotype; D051381:Rats; D013234:Stem Cells / Q000145:classification / Q000166:cytology* / Q000276:immunology; D006377:T-Lymphocytes, Helper-Inducer / Q000276:immunology; D013997:Time Factors
D003911:Dextrans; D008070:Lipopolysaccharides; D016264:Dextran Sulfate