论文部分内容阅读
目的：建立胚胎干细胞体外定向诱导分化为浆细胞的体系，为研究造血发生的基因调控及Ｂ淋巴细胞发育过程奠定基础。方法：分阶段培养，第一、二阶段使用半固体培养，分组加入细胞因子、２－巯基乙醇和胚胎细胞条件培养液，用流式细胞仪、免疫组化及Ｗｒｉｇｈｔ’ｓ染色鉴定细胞，第三阶段移入骨髓基质细胞滋养层。结果：第一阶段：加入ＳＣＦ、ＴＰＯ、ＩＬ－６、ＩＬ－３、Ｅｐｏ、和Ｇ－ＣＳＦ，４ｄ后细胞大片坏死，仅加入２－巯基乙醇可形成胚胎体。第二阶段：加入ＳＣＦ和ＴＰＯ，细胞增殖缓慢，再加入条件培养液，发育３～４ｄ可形成ＣＤ３４＋的细胞集落。第三阶段：移至骨髓基质细胞饲养层后，发育为浆细胞。结论：胚胎干细胞在体外可定向分化为浆细胞。胚胎干细胞在早期发育不需要细胞因子，胚胎细胞的条件培养液可促使其向造血细胞发育，骨髓基质细胞饲养层可使其向浆细胞分化
OBJECTIVE: To establish a system in which ESCs differentiate into plasma cells in vitro, and lay the foundation for the study on the gene regulation of hematopoiesis and the development of B lymphocytes. Methods: The cells were cultured in stages. In the first and second stages, semi-solid culture was used. Cytokines, 2-mercaptoethanol and embryonic cells conditioned medium were added in groups. The cells were identified by flow cytometry, immunohistochemistry and Wright’s staining. Three stages into the marrow stromal cells trophoblast. Results: In the first phase, after 4 days, SCF, TPO, IL-6, IL-3, Epo and G-CSF were added. The second stage: adding SCF and TPO, cell proliferation is slow, then add the conditioned medium, the development of 3 ~ 4d can form CD34 + cell colonies. The third stage: moved to the marrow stromal cells feeder layer, the development of plasma cells. Conclusion: Embryonic stem cells can be differentiated into plasma cells in vitro. Embryonic stem cells in the early development does not require cytokines, embryonic cells conditioned medium can promote its development to hematopoietic cells, bone marrow stromal cell feeder layer can make it to plasma cells