Precursor Lymphoblastic Lymphoma / Acute Lymphoblastic Leukemia Cells

Object: To investigate method of isolation and identification of acute lymphoblastic leukemic stem cells in a T cell lymphoblastic leukemic cell line Jurkat as well as primary acute lymphoblastic leukemic cells.Methods: Immunophenotypes of Jurkat cells were detected by Flowcytometry. Cells expressed CD34 was positively selected by using a magnetically actived cell sorting kit, CD34- cells were acquired by monoclone cell culture. Levels of expression of CD34 and stem cell associated genes were compared by RT-PCR for CD34+ and CD34- monoclones. Clonogenesis ability of the CD34+ and CD34- cells were tested by soft agarose assay and in vivo tumorigenesis abilities were tested in severe combined immunodeficient (SCID) mice. Primary acute lymphoblastic leukemic cells of different differentiation stages were purified by fluorescence-activated cell sorting. Common chromosome-translocation types of acute lymphoblastic leukemia were detected by RT-PCR..Results: All Jurkat cells expressed committed T cell lineage markers. Jurkat cells could be divided into two differentiated cell groups by CD34 expression status, which were CD34+CD38+CD7+ and CD34-CD38+CD7+ cells. Both in vitro and in vivo experiments proved that part of CD34+ cells could differentiate into CD34- cells by continuously decreasing of CD34 gene expression , this process was not reversible. Both CD34+ and CD34- monoclone cells were capable of extensive proliferation. Expression of hematopoietic stem cell associated genes like Wnt1, Bmi1, Lefl, ABCG2 and TERT showed no significant differences between CD34+ and CD34- monoclones. Results of soft agorose monoclone formation experiment and in vivo SCID mice tumorigenesis experiment showed no significant differences between CD34+ and CD34- monoclone cells.Conclusion: Positively selected CD34 megnetically activated cell sorting method is effective in isolating CD34+ cells. All of Jurkat cells express T cell-lineage markers, which may suggest that Jurkat cell line originated from T-cell committed progenitor cells characterized by...