两种人羊膜细胞的表型鉴定和分化潜能测定(英文)

Phenotypic Identification and Differentiation Potential Analysis of Two Kinds of Human Amniotic Cells

本研究旨在分离、培养和表型鉴定两种人羊膜细胞并分析其多向分化潜能。羊膜中可分离出来源于外胚层的羊膜上皮细胞和来源于中胚层的羊膜间充质细胞。用流式细胞术和免疫荧光法对其进行表型鉴定,同时用免疫荧光法分析其多向分化潜能。结果表明:两种人羊膜细胞阳性表达HLA-A,B,C和间充质干细胞的标志(CD29,CD73,CD44,CD59,CD90,CD105,CH166),不表达造血干细胞的标志(CD31,CD34,CD45,HLA-DR),弱表达协同刺激分子(CD40,CD40L,CD80,CD86),且这些表型的表达量在第3-7代都维持稳定。免疫荧光法显示羊膜上皮细胞表达角蛋白19,不表达波形蛋白,而羊膜间充质细胞则相反。对其多向分化潜能测定显示,羊膜间充质细胞更好地向心肌细胞分化,而羊膜上皮细胞更好地向神经细胞分化。结论:从羊膜中可分离出羊膜上皮细胞和羊膜间充质细胞,两种细胞的表型相似,多向分化潜能却不同。羊膜上皮细胞具有更好的外胚层分化潜能,而羊膜间充质细胞更好地向中胚层分化。此结论对细胞治疗有很好的指导作用。

The aim of this study was to isolate, cultivate and phenotypicaUy characterize two types of human amnio- tic membrane （HAM）-derived cells, and to analyze their differentiation potential in vitro. Human amnion epithelial cells （hAEC） were derived from the embryonic ectoderm, while human amnion mesenchymal cells （hAMC） were derived from the embryonic mesoderm. The cells were characterized by flow cytometry and immunofluorescence, then immuno- fluorescence also was performed for the analysis of multipotentiality in differentiation. The results indicated that immuno- phenotypic characterization of both cell types demonstrated positive for HLA-A, B, C and mesenchymal stem cell markers （CD29, CD73, CD44, CD59, CDg0, CD105, CD166）, but did not express the hematopoietic markers （CD31, CD34, CIMS, HLA-DR） and showed the weak expression of costimulatory molecules（CD40, CD40L, CD80, CD86）. Phenotypes of both cell populations were maintained from passages 3 to 7. The immunofluorescence indicated that hAEC expressed cytokeratin 19, but did not express vimentin. On the contrary, hAMC expressed vimentin but did not express cytokeratin 19. The assessment of multilineage potential demonstrated that hAMC showed greater cardiomyocytic poten- tial, while hAEC showed greater neural potential. It is concluded that hAEC and hAMC can be successfully isolated from the HAM. Both cell populations possess similar immunophenotype. However, they differ in cell yield and multipotential for differentiation into the major lineages, hAEC possess a much greater ectodermal differentiation capacity, while hAMC possess a much greater mesodermal differentiation capacity. This conclusion will be important for use of these cells in cell therapy.