Dynamics of donor cell persistence and recipient cell replacement in orthotopic corneal allografts in mice.
Dynamics of donor cell persistence and recipient cell replacement in orthotopic corneal allografts in mice.
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发表时间：
2001-07
影响因子：
4.4
通讯作者：
J. Hori;J. Streilein
中科院分区：
文献类型：
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作者：
J. Hori;J. Streilein
目的确定供体细胞在小鼠原位角膜移植物的三层细胞中的存留和受体细胞在三层细胞中的再填充的程度。方法以BALB/c、C57 BL/6和增强型绿色荧光蛋白（EGFP）转基因小鼠（B6背景）为供体和受体，进行同基因和同种异体角膜移植。分别于术后第5、10、15、28、56天取移植眼，碘化丙啶染色，共聚焦显微镜逐层观察。移植物中的骨髓源性细胞用免疫化学方法进行评估。结果在同基因和同种异体移植物中，供体上皮在15天内完全被受体上皮细胞取代，而供体基质角膜细胞和内皮细胞在同基因移植物和接受的同种异体移植物中几乎保持完整。在被排斥的同种异体移植物中，既没有检测到供体来源的角膜细胞，也没有检测到内皮细胞，相反，大量存在供体来源的基质成纤维细胞、新血管和浸润性白细胞。排斥移植物的后表面缺乏角膜内皮，并且不完全被受体来源的骨髓源性细胞覆盖。结论：在小鼠中，移植物来源的上皮细胞与角膜移植结果基本无关，而供体来源的内皮细胞和角膜细胞的持久性与移植物的接受性完全相关。角膜内皮不能覆盖接受或排斥的角膜移植物的后表面，而受体来源的骨髓来源的细胞在排斥的移植物中占据该部位。
PURPOSE To determine the extent to which donor cells persist and recipient cells repopulate each of the three cell layers of orthotopic corneal grafts in mice. METHODS BALB/c, C57BL/6, and enhanced green fluorescence protein (EGFP) transgenic mice (B6 background) were used as donors and recipients for orthotopic syngeneic and allogeneic corneal grafts. Graft-bearing eyes were harvested at 5, 10, 15, 28, and 56 days, stained with propidium iodide, and observed (layer by layer) by confocal microscopy. Bone marrow-derived cells in the grafts were assessed immunohistochemically. RESULTS Donor epithelium was totally replaced by recipient epithelial cells within 15 days in both syngeneic and allogeneic grafts, whereas donor stromal keratocytes and endothelial cells were retained virtually intact in syngeneic grafts and in accepted allografts. In rejected allografts, neither donor-derived keratocytes nor endothelial cells were detected, and, instead, recipient-derived stromal fibroblasts, neovessels, and infiltrating leukocytes were heavily represented. The posterior surface of rejected grafts was devoid of corneal endothelium and was covered incompletely with bone marrow-derived cells of recipient origin. CONCLUSIONS Whereas in mice graft-derived epithelium is largely irrelevant to corneal allograft outcome, persistence of donor-derived endothelium and keratocytes correlates perfectly with graft acceptance. Recipient endothelium is incapable of covering the posterior surface of accepted or rejected corneal grafts, whereas bone marrow-derived cells of recipient origin come to occupy this site in rejected grafts.