Functional characterization of B lymphocytes generated in vitro from embryonic stem cells.
To study molecular events involved in B lymphocyte development and V(D)J rearrangement, we have established an efficient system for the differentiation of embryonic stem (ES) cells into mature Ig-secreting B lymphocytes. Here, we show that B lineage cells generated in vitro from ES cells are functionally analogous to normal fetal liver-derived or bone marrow-derived B lineage cells at three important developmental stages: first, they respond to Flt-3 ligand during an early lymphopoietic progenitor stage; second, they become targets for Abelson murine leukemia virus (A-MuLV) infection at a pre-B cell stage; third, they secrete Ig upon stimulation with lipopolysaccharide at a mature mitogen-responsive stage. Moreover, the ES cell-derived A-MuLV-transformed pre-B (EAB) cells are phenotypically and functionally indistinguishable from standard A-MuLV-transformed pre-B cells derived from infection of mouse fetal liver or bone marrow. Notably, EAB cells possess functional V(D)J recombinase activity. In particular, the generation of A-MuLV transformants from ES cells will provide an advantageous system to investigate genetic modifications that will help to elucidate molecular mechanisms in V(D)J recombination and in A-MuLV-mediated transformation.
Cho
S K
SK
Department of Immunology, University of Toronto, Toronto, ON M5S 1A8, Canada.
Webber
T D
TD
Carlyle
J R
JR
Nakano
T
T
Lewis
S M
SM
Zúñiga-Pflücker
J C
JC
D000011:Abelson murine leukemia virus; D000818:Animals; D001402:B-Lymphocytes / Q000502:physiology*; D002454:Cell Differentiation / Q000187:drug effects; D018920:Coculture Techniques; D015322:Gene Rearrangement, B-Lymphocyte; D006410:Hematopoiesis; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000502:physiology*; D008024:Ligands; D008565:Membrane Proteins / Q000494:pharmacology; D051379:Mice; D016133:Polymerase Chain Reaction
D008024:Ligands; D008565:Membrane Proteins; C084881:flt3 ligand protein