Chen Hung-Chi Jesse, Chen Hsiang-Ling, Lai Jui-Yang, Chen Chih-Chun, Tsai Yueh-Ju, Kuo Ming-Tse, Chu Pao-Hsien, Sun Chi-Chin, Chen Jan-Kan, Ma David Hui-Kang
Limbal Stem Cell Laboratory, Department of Ophthalmology, Chang Gung Memorial Hospital, Taoyuan, Taiwan.
Invest Ophthalmol Vis Sci. 2009 Oct;50(10):4660-8. doi: 10.1167/iovs.09-3377. Epub 2009 May 20.
To determine the expression of differentiation and progenitor cell markers in corneal tissues that previously underwent autologous cultivated oral mucosal epithelial transplantation (COMET).
Four eyes from three alkaline-injured patients and one thermally injured patient underwent COMET to promote re-epithelialization or corneal reconstruction. Between 10 and 22 months (mean, 14.2 +/- 5.5 months [SD]) after COMET, the corneal tissues were obtained after penetrating keratoplasty (n = 1) or autologous limbal transplantation (n = 3). Immunoconfocal microscopy for keratin (K)3, -12, -4, -13, and -8; connexin (Cx)43; MUC5AC; laminin-5; pan-p63; ABCG2; and p75 was performed in those specimens as well as in the oral mucosa and cultivated oral mucosal epithelial cells (OMECs).
All four specimens were unanimously positive for K3, -4, and -13 but negative for K8 and MUC5AC, suggesting that the keratinocytes were oral mucosa-derived. However, peripheral K12 staining was positive only in patient 2, suggesting a mixed oral and corneal epithelium in that case. Cx43 staining in the basal epithelium was negative in patients 1, 2, and 3, but was positive in patient 4. Small, compact keratinocytes in the basal epithelium preferentially expressed pan-p63, ABCG2, and p75. Although the staining of pan-p63 and ABCG2 tended to be more than one layer, signal for p75 was consistently localized only to the basal layer.
The study demonstrated the persistence of transplanted OMECs in human corneas. In addition, small, compact cells in the basal epithelium preferentially expressed the keratinocyte stem/progenitor cell markers, which may be indicative of the engraftment of the progenitor cells after transplantation.
确定先前接受自体培养口腔黏膜上皮移植（COMET）的角膜组织中分化和祖细胞标志物的表达情况。
对3例碱烧伤患者和1例热烧伤患者的4只眼进行COMET，以促进上皮再形成或角膜重建。在COMET后10至22个月（平均14.2±5.5个月[标准差]），通过穿透性角膜移植术（n = 1）或自体角膜缘移植术（n = 3）获取角膜组织。对这些标本以及口腔黏膜和培养的口腔黏膜上皮细胞（OMEC）进行角蛋白（K）3、-12、-4、-13和-8、连接蛋白（Cx）43、MUC5AC、层粘连蛋白-5、泛p63、ABCG2和p75的免疫共聚焦显微镜检查。
所有4个标本的K3、-4和-13均呈一致阳性，但K8和MUC5AC呈阴性，提示角质形成细胞来源于口腔黏膜。然而，仅患者2的周边K12染色呈阳性，提示该病例中存在口腔和角膜上皮混合的情况。患者1、2和3的基底上皮Cx43染色呈阴性，但患者4呈阳性。基底上皮中的小而致密的角质形成细胞优先表达泛p63、ABCG2和p75。尽管泛p63和ABCG2的染色往往不止一层，但p75信号始终仅局限于基底层。
该研究证明了移植的OMEC在人角膜中的持续存在。此外，基底上皮中的小而致密的细胞优先表达角质形成干细胞/祖细胞标志物，这可能表明移植后祖细胞的植入。
