[摘要] 目的：研究与羊膜上皮细胞（amniotic epithelial cells，AEC）共培养对羊膜间充质干细胞（amniotic mesenchymal stem cells，AMSC）生物学特性的影响，并探讨基质细胞衍生因子1(stromal cell derived factor-1, SDF-1）/CXCR4 轴在AMSC 归巢、迁移过程中的作用。方法：从人羊膜中分别分离、培养AMSC和AEC，进行扩增及鉴定。实验设AMSC与AEC共培养组、无血清AMSC培养组以及血清AMSC培养组，培养24、48、72 h 后，通过CCK-8 及锥虫蓝实验检测AMSC细胞增殖活性，免疫荧光流式细胞术和Real-time PCR检测CXCR4 mRNA的表达水平，细胞迁移实验检测AMSC细胞体外迁移能力。结果：48、72 h 共培养组及血清培养组细胞增殖活性及增殖率均高于无血清培养组(P<0.05)；共培养组与无血清培养组AMSC细胞的CXCR4 mRNA和蛋白表达明显高于血清培养组(P<0.05)，两组AMSC细胞依赖迁移能力明显高于血清培养组(P<0.05)。结论：与AEC共培养的AMSC细胞仍具备间充质干细胞的基本生物学特性，且能保持良好的增殖活性，共培养能上调AMSC表面CXCR4，并在体外增强其迁移能力。

[Abstract] Objective: The aim of this study was to investigate the effect of co-culture with AEC (amniotic epithelial cell) on the biological characteristics of AMSC (amniotic mesenchymal stem cell), and to investigate the roles of SDF-1/CXCR4 axis in the homing and migration of AMSC. Methods: AMSC and AEC were isolated from human amnion, and then cultured, amplified and identified, respectively.The AMSC were divided into three groups: AEC co-cultured group, serum-free cultured group and serum cultured group. After culture for 24 h, 48 h, and 72 h, the proliferation viability of AMSC was measured by CCK-8 assay and trypan blue staining; the expression of CXCR4 mRNA was analyzed by flow cytometry and Real-time RT-PCR, and the migration ability of AMSC in vitro was observed by migration assay. Results: Cell viability (48 h and 72 h) and survival rate in the co-culture and serum groups were higher than those in the serum-free group (all P<0.05). The mRNA and protein expressions of CXCR4 in AMSC of the co-culture and serum-free groups were significantly higher than those of the serum group (P<0.05). The migration ability of AMSC in the co-culture and serumfree groups, which increase with the SDF-1 (stromal cell derived factor-1) concentration gradient, were higher than that in the serum group (P<0.05). Conclusion: AMSC co-cultured with AEC still have the basic biological characteristics of MSC, and showed good growth activity. Co-culture with AEC can up-regulate CXCR4 on AMSC surfaces and enhance the migration ability of AMSC in vitro.

国家自然科学基金资助项目（No. 31260232）；云南省医疗卫生单位内设研究机构科研项目（No.2016NS048）；云南省科技厅-昆明医科大学应用基础研究联合专项基金资助项目（No. 2017FE468-030）