中文摘要：

目的通过组织块培养法,建立一种高效可靠的小鼠脾脏间充质干细胞分离扩增策略。方法无菌条件下取小鼠脾脏组织充分碾碎,使用胶原酶消化获得的脾脏基质组织,将消化后的组织块种于培养瓶中,观察从组织块中迁移出的细胞形态,采用流式细胞术分析其表型,并将其做成骨和成脂肪诱导分化。结果消化后的脾脏组织块中迁移迁移出大量梭形纤维样细胞,流式细胞术检测结果表明,该种细胞高表达CD29、CD44、Sca-1和CD105,低表达CD11b、CD34、CD45和Ia。多向分化实验结果表明：小鼠脾脏基质组织中迁移出的细胞具有较好的成脂肪和成骨分化能力。结论组织块法可以分离培养出小鼠脾脏间充质干细胞,所获得的细胞纯度高,分化能力强,为开展相关研究提供了良好的细胞模型。

英文摘要：

Objective This study aimed to establish a reliable primary culture protocol for preparing murine spleen-derived mesenchymal stem cells（ MSCs） by tissue explant culture. Methods Healthy mouse spleens were crushed by syringe handle to harvest spleen mesenchymal tissues. Then the tiny pieces of spleen tissue were digested by collagenase II before seeded into culture flasks. The morphological characteristics of spleen tissue-derived cells were observed under the inverted microscope. Further,the surface antigen profile of the cells was analyzed by flow cytometry（ FACS）. The cells were induced to differentiate into osteoblasts and adipocytes. Results The murine spleen-derived MSCs exhibited a spindle-shaped appearance. The FACS results showed that the spleen-derived MSCs highly expressed CD29,CD44,CD105 and Sca-1, but weakly expressed CD11 b, CD34, CD45 and Ia. In addition, the spleen-derived MSCs steadily differentiated into osteoblasts and adipocytes in the induction medium. Conclusions A method of primary culture of murine spleen-derived MSCs by explant culture is successfully established. The harvested MSCs exhibit high purity and cell proliferation ability,and provide a reliable cell model for related researches.