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人未成熟牙髓干细胞与角膜缘干细胞具有关键特征。

Human immature dental pulp stem cells share key characteristic features with limbal stem cells.

作者信息

Monteiro B G, Serafim R C, Melo G B, Silva M C P, Lizier N F, Maranduba C M C, Smith R L, Kerkis A, Cerruti H, Gomes J A P, Kerkis I

机构信息

Genetics Laboratory, Butantan Institute, São Paulo, SP, Brazil.

出版信息

Cell Prolif. 2009 Oct;42(5):587-94. doi: 10.1111/j.1365-2184.2009.00623.x. Epub 2009 Jul 14.

Abstract

OBJECTIVES

Limbal stem cells (LSC) are self-renewing, highly proliferative cells in vitro, which express a set of specific markers and in vivo have the capacity to reconstruct the entire corneal epithelium in cases of ocular surface injury. Currently, LSC transplantation is a commonly used procedure in patients with either uni- or bilateral total limbal stem cells deficiency (TLSCD). Although LSC transplantation holds great promise for patients, several problems need to be overcome. In order to find an alternative source of cells that can partially substitute LSC in cornea epithelium reconstruction, we aimed at investigating whether human immature dental pulp stem cells (hIDPSC) would present similar key characteristics as LSC and whether they could be used for corneal surface reconstruction in a rabbit TLSCD model.

MATERIALS

We used hIDPSC, which co-express mesenchymal and embryonic stem cell markers and present the capacity to differentiate into derivative cells of the three germinal layers. TLSCD was induced by chemical burn in one eye of rabbits. After 30 days, the opaque tissue formed was removed by superficial keratectomy. Experimental group received undifferentiated hIDPSC, while control group only received amniotic membrane (AM). Both groups were sacrificed after 3 months.

RESULTS AND CONCLUSIONS

We have demonstrated, using immunohistochemistry and reverse transcription-polymerase chain reaction, that hIDPSCs express markers in common with LSC, such as ABCG2, integrin beta1, vimentin, p63, connexin 43 and cytokeratins 3/12. They were also capable of reconstructing the eye surface after induction of unilateral TLSCD in rabbits, as shown by morphological and immunohistochemical analysis using human-specific antibodies against limbal and corneal epithelium. Our data suggest that hIDPSCs share similar characteristics with LSC and might be used as a potential alternative source of cells for corneal reconstruction.

摘要

目的

角膜缘干细胞(LSC)在体外具有自我更新和高度增殖能力,表达一组特定标志物,在体内,眼表损伤时能够重建整个角膜上皮。目前,LSC移植是单侧或双侧完全角膜缘干细胞缺乏(TLSCD)患者常用的治疗方法。尽管LSC移植给患者带来了很大希望,但仍有几个问题需要克服。为了找到一种能够部分替代LSC进行角膜上皮重建的细胞替代来源,我们旨在研究人未成熟牙髓干细胞(hIDPSC)是否具有与LSC相似的关键特性,以及它们是否可用于兔TLSCD模型的角膜表面重建。

材料

我们使用了共表达间充质和胚胎干细胞标志物且具有分化为三个胚层衍生细胞能力的hIDPSC。通过化学烧伤诱导兔的一只眼睛发生TLSCD。30天后,通过浅表角膜切除术切除形成的不透明组织。实验组接受未分化的hIDPSC,而对照组仅接受羊膜(AM)。3个月后处死两组动物。

结果与结论

我们通过免疫组织化学和逆转录 - 聚合酶链反应证明,hIDPSC表达与LSC共同的标志物,如ABCG2、整合素β1、波形蛋白、p63、连接蛋白43和细胞角蛋白3/12。使用针对角膜缘和角膜上皮的人特异性抗体进行形态学和免疫组织化学分析表明,它们在诱导兔单侧TLSCD后也能够重建眼表。我们的数据表明,hIDPSC与LSC具有相似的特性,可能作为角膜重建的潜在细胞替代来源。

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