CAJ | 학술논문

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.
본연구지재분리、배양화표형감정량충인양막세포병분석기다향분화잠능.양막중가분리출래원우외배층적양막상피세포화래원우중배층적양막간충질세포.용류식세포술화면역형광법대기진행표형감정,동시용면역형광법분석기다향분화잠능.결과표명:량충인양막세포양성표체HLA-A,B,C화간충질간세포적표지( CD29,CD73,CD44,CD59,CD90,CD105,CH166),불표체조혈간세포적표지(CD31,CD34,CD45,HLA-DR),약표체협동자격분자( CD40,CD40L,CD80,CD86),차저사표형적표체량재제3-7대도유지은정.면역형광법현시양막상피세포표체각단백19,불표체파형단백,이양막간충질세포칙상반.대기다향분화잠능측정현시,양막간충질세포경호지향심기세포분화,이양막상피세포경호지향신경세포분화.결론:종양막중가분리출양막상피세포화양막간충질세포,량충세포적표형상사,다향분화잠능각불동.양막상피세포구유경호적외배층분화잠능,이양막간충질세포경호지향중배층분화.차결론대세포치료유흔호적지도작용.
The aim of this study was to isolate,cultivate and phenotypically characterize two types of human amniotic 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 immunofluorescence also was performed for the analysis of multipotentiality in differentiation.The results indicated that immunophenotypic characterization of both cell types demonstrated positive for HLA-A,B,C and mesenchymal stem cell markers (CD29,CD73,CD44,CD59,CD90,CD105,CD166),but did not express the hematopoietic markers (CD31,CD34,CD45,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 potential,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.