Human bone marrow CD34~+ cells differentiate to T cells in thymic and bone marrow stromal microenvironment in vitro / 中国免疫学杂志

ABSTRACT

Objective:To study methods for inducing CD34+ cells of human bone marrow to differentiate into T cells in vitro to provide theory and method basics for the investigation of activity of T cells derived from psoriatic bone marrow CD34+ cells and establish a technological platform to investigate T lymphopoiesis activity of hematopoietic cells.Methods:Bone marrow CD34+ hematopoietic progenitor cells were isolated by immunomagnetic cell selection and induced differentiate into T cells in the bone in the marrow and thymic stromal microenvironment.Immunfluorescence dying method and flow cytometry analysis were performed to detect CD1-CD3+ cells,CD3+CD4+CD8-cells and CD3+CD4-CD8+ cells dynamically.Results:In the first week,the non-adhension cells were composed mostly of immature CD1+CD3-cells and CD1+CD3+ cells and small proportion of mature CD1-CD3+ cells.In the following analysis,the proportion of immature cells rapidly decreased and CD1-CD3+ cells increased.After 1 week culture,CD4+CD8+ double positive T cells and a small population CD4+CD8-and CD4-CD8+ could be detected among the CD3+ cells.In the following culture,the proportion of CD4+CD8+ double positive T cells decreased significantly and single positive T cells increased gradually.However,small proportion of mature T cells could be detected in the early stage and cann't be found after 4 weeks in the culture system without thymic stromal cells.Conclusion:Mature single positive T cells can develop from CD34+ hematopoietic progenitor cells in the bone marrow and thymic stromal microenvironment and the thymic stromal cells are vital for T lymphopoiesis.