中文摘要：

目的：研究人骨髓CD34^＋细胞体外向T细胞定向分化的方法，为研究造血细胞淋系造血活性及T细胞发育和分化提供技术平台。方法：免疫磁珠法分离骨髓CD34^＋细胞，在骨髓基质细胞条件培养液构建的微环境下，在胸腺基质细胞的支持下，使其体外向T细胞定向分化，收集培养的非贴壁细胞，免疫荧光染色后经流式细胞术检测培养不同时间CD1^-CD3^＋细胞、CD3^＋CD4^＋CD8^-细胞及CD3^＋CD4^-CD8^＋细胞比例。结果：培养1周时，培养细胞中以不成熟的CD1^＋CD3^-细胞、CD1^＋CD3^＋细胞为主，可检测到少量CD1^-CD3^＋细胞，随培养时间延长，不成熟细胞比例逐渐减少，而成熟的CD1^-CD3^＋细胞比例逐渐增加；在CD3^＋细胞中，培养初期以不成熟的双阳性细胞CD4^＋CD8^＋为主，而成熟的单阳性CD^＋CD8^-细胞及CD4^-CD8^＋细胞占极小比例，随培养时间延长，双阳性细胞比例逐渐减少，而成熟的单阳性细胞比例逐渐增高；而无胸腺基质细胞支持的CD34＋细胞仅在培养初期检测到成熟细胞存在，而培养后4周基本检测不到成熟T细胞的存在。结论：在骨髓基质细胞及胸腺基质细胞的支持下，骨髓CD34^＋细胞可体外发育为成熟的CD1^-CD3^＋细胞及单阳性T细胞，其中胸腺基质细胞的支持对于造血细胞向T细胞的体外定向分化极其重要。

英文摘要：

Objective： To study methods for inducing CD3^4＋ 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 teehnological 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 Iramunfluorescence dying method and flow cytometry analysis were performed to detect CD1^- CD3^＋ cells, CD3^＋ CD4^＋ CD8^- cells and CD3^＋ CD^-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, CIM ＋ 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 CD^＋ CD8^＋ double positive T cells decreased significandy and single positive T cells increased gradually. However, small proportion of mature T ceils 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 and the thymic stromal cells are vital for T lymphopoiesis.