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Microenvironment effect of APA microcapsule on embryonic stem cell proliferation and differentiation
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- Author:
- No author available
- Journal Title:
- ACTA PHYSIOLOGICA SINICA
- Issue:
- 6
- DOI:
- 10.3321/j.issn:0371-0874.2005.06.016
- Key Word:
- 胚胎干细胞;微胶囊;微环境;分化
Abstract： 以小鼠胚胎干细胞(embryonic stem cell,ESC)为模型,在生理条件下对ESC进行微囊化包封、培养,并利用免疫组织化学技术及RT-PCR方法检测其生长及未分化状态,以期建立微囊化ESC这一体外培养模型,同时明确海藻酸钠-聚赖氨酸-海藻酸钠(alginate-poly-lysine-alginate,APA)微囊微环境对ESC增殖及分化潜能的影响.结果表明:ESC能够在微囊(包括液化型及非液化型)或微球(海藻酸钙胶珠)内生长良好,但因生长环境存在差异,其表现的生长行为各具特征.比较其它类型,ESC在液化型APA微囊内的存活期限最长.经体外维持培养3周以上,仍能持续表达胚胎源未分化干细胞的标志性蛋白AP,SSEA-1及转录因子Oct-4.为进一步明确微囊内增殖的ESC是否仍具有多向分化的干细胞潜能,应用机械破囊法释放微囊内ESC团,并在体外进行定向诱导.经过近3周的条件诱导,其结果为:细胞团DTZ染色阳性;anti-insulin免疫荧光检测阳性;且特异性表达Pdx-1,Ins-1基因.上述结果证明:APA微囊为ESC维持未分化状态的增殖提供了特殊的微环境,APA微囊内所形成的ESC团仍具有多向分化的干细胞潜能.
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