Harkin D G, Barnard Z, Gillies P, Ainscough S L, Apel A J G
Tissue BioRegeneration and Integration Program, Institute for Health and Biomedical Innovation, Queensland University of Technology, 2 George Street, Brisbane 4000, Australia.
Br J Ophthalmol. 2004 Sep;88(9):1154-8. doi: 10.1136/bjo.2003.037853.
To investigate the expression of p63 and cytokeratins throughout the course of producing a cultivated autograft of limbal epithelial cells.
A 75 year old male with a severe alkali burn to his right eye received two cultivated autografts of limbal epithelial cells on amniotic membrane followed by a corneal allograft. Immunostaining for p63 and cytokeratins was performed during ex vivo expansion with 3T3 fibroblasts, following subcultivation on amniotic membrane, and on the excised corneal button.
Cultures grown in the presence of 3T3 fibroblasts or on amniotic membrane displayed positive staining for keratins 14 and 19, and p63, but poor staining for keratin 3 (K3). The excised corneal button possessed a stratified epithelium of K3 positive cells residing on amniotic membrane.
Our results document for the first time the co-expression of cytokeratins 14 and 19 with p63 in a cultivated limbal graft. These data support the conclusion that cultivated grafts of limbal epithelium contain predominantly undifferentiated cells with the potential to regenerate a normal corneal epithelium.
研究在制备培养的自体角膜缘上皮细胞移植片的整个过程中p63和细胞角蛋白的表达情况。
一名75岁右眼遭受严重碱烧伤的男性接受了两片培养的羊膜上自体角膜缘上皮细胞移植片，随后进行了同种异体角膜移植。在用3T3成纤维细胞进行体外扩增期间、在羊膜上再次培养后以及在切除的角膜植片上，对p63和细胞角蛋白进行免疫染色。
在3T3成纤维细胞存在下或在羊膜上培养的细胞对角蛋白14和19以及p63呈阳性染色，但对角蛋白3（K3）染色较差。切除的角膜植片在羊膜上有一层K3阳性细胞的复层上皮。
我们的结果首次证明了在培养的角膜缘移植物中细胞角蛋白14和19与p63的共表达。这些数据支持这样的结论，即培养的角膜缘上皮移植物主要包含具有再生正常角膜上皮潜力的未分化细胞。
