Amnion epithelial cell isolation and characterization for clinical use.
Amnion epithelial cell isolation and characterization for clinical use.
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DOI：
10.1002/9780470151808.sc01e06s13
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发表时间：
2010-04-01
影响因子：
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通讯作者：
Jenkin, Graham
中科院分区：
文献类型：
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作者：
Murphy, Sean;Rosli, Sharina;Jenkin, Graham
人羊膜上皮细胞 (hAEC) 是干细胞标记物呈阳性的异源细胞群；它们表现出多谱系分化潜力，分化为内胚层（肝、肺上皮）、中胚层（骨、脂肪）和外胚层（神经细胞）细胞。它们具有低免疫原性并具有有效的免疫抑制特性。因此，hAEC可能是细胞治疗的宝贵细胞来源。本单元描述了一种高效且有效的 hAEC 分离、培养和冷冻保存方法，该方法不含动物产品，并且符合当前临床使用细胞制备指南。使用该方法分离的细胞在传代 5 代后通过分析核型、细胞周期分布和端粒长度变化进行表征。通过分化为三个初级胚层的谱系以及通过 PCR、免疫细胞化学和组织学分析的谱系特异性标记物的表达，证明了使用这种无动物产品的方法分离的 hAEC 的分化潜力。
Human amnion epithelial cells (hAECs) are a heterologous population positive for stem cell markers; they display multilineage differentiation potential, differentiating into cells of the endoderm (liver, lung epithelium), mesoderm (bone, fat), and ectoderm (neural cells). They have a low immunogenic profile and possess potent immunosuppressive properties. Hence, hAECs may be a valuable source of cells for cell therapy. This unit describes an efficient and effective method of hAEC isolation, culture, and cryopreservation that is animal product-free and in accordance with current guidelines on preparation of cells for clinical use. Cells isolated using this method were characterized after 5 passages by analysis of karyotype, cell cycle distribution, and changes in telomere length. The differentiation potential of hAECs isolated using this animal product-free method was demonstrated by differentiation into lineages of the three primary germ layers and expression of lineage-specific markers analyzed by PCR, immunocytochemistry, and histology.