Survival of human embryonic stem cells implanted in the guinea pig auditory epithelium.
Survival of human embryonic stem cells implanted in the guinea pig auditory epithelium.
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DOI：
10.1038/srep46058
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发表时间：
2017-04-07
影响因子：
4.6
通讯作者：
Raphael Y
中科院分区：
文献类型：
--
作者：
Lee MY;Hackelberg S;Green KL;Lunghamer KG;Kurioka T;Loomis BR;Swiderski DL;Duncan RK;Raphael Y
成熟耳蜗中的毛细胞不能自发再生。干细胞疗法是修复毛细胞的一种潜在方法。然而，当细胞被移植到耳蜗的中阶（SM）时，它们由于高钾浓度而迅速死亡。我们之前描述了一种调节SM的方法，使其更适合植入细胞，并表明HeLa细胞可以使用这种方法存活长达一周。在这里，我们评估了组成型表达GFP（H9 Cre-LoxP）的人胚胎干细胞（hESC）在经过预处理以降低钾水平的聋豚鼠耳蜗中的存活情况。注射后至少7天，耳蜗中可检测到GFP阳性细胞。细胞呈球形或不规则形，部分细胞聚集。在移植前用癸酸钠冲洗SM导致表达多能性标志物Oct 3/4的干细胞比例降低，并增加细胞存活。数据表明，旨在瞬时降低SM中钾浓度的调节程序促进hESC存活至少一周。在该时间窗期间，可以应用额外的程序来启动植入的hESC分化成新的毛细胞。
Hair cells in the mature cochlea cannot spontaneously regenerate. One potential approach for restoring hair cells is stem cell therapy. However, when cells are transplanted into scala media (SM) of the cochlea, they promptly die due to the high potassium concentration. We previously described a method for conditioning the SM to make it more hospitable to implanted cells and showed that HeLa cells could survive for up to a week using this method. Here, we evaluated the survival of human embryonic stem cells (hESC) constitutively expressing GFP (H9 Cre-LoxP) in deaf guinea pig cochleae that were pre-conditioned to reduce potassium levels. GFP-positive cells could be detected in the cochlea for at least 7 days after the injection. The cells appeared spherical or irregularly shaped, and some were aggregated. Flushing SM with sodium caprate prior to transplantation resulted in a lower proportion of stem cells expressing the pluripotency marker Oct3/4 and increased cell survival. The data demonstrate that conditioning procedures aimed at transiently reducing the concentration of potassium in the SM facilitate survival of hESCs for at least one week. During this time window, additional procedures can be applied to initiate the differentiation of the implanted hESCs into new hair cells.