Department of Ophthalmology, University Hospital Antwerp, Antwerp, Belgium.
Tissue Eng Part C Methods. 2010 Oct;16(5):921-7. doi: 10.1089/ten.TEC.2009.0634.
To describe a standardized, xenogenic-free protocol for the manufacture of limbal epithelial stem cell grafts and a "no touch" surgical technique for its standardized transplantation.
Antwerp University Hospital, Antwerp, Belgium.
The limbo-amnion composite graft is generated by cultivating limbal epithelial stem cells on a standardized (thermolysin treated and spongy layer removed) amniotic membrane, stretched within an interlockable amnion ring. The cells are cultured in CnT-20 medium with the addition of 1% human AB serum for a period of 2 weeks. Fibrin glue is applied to the surgically prepared recipient's cornea and in one fluid motion, the composite graft within the amnion ring construct is transferred from culture and positioned onto the graft bed. The required size is cut out at the level of the limbus by means of a trephine and/or microsurgical scissors.
The lightweight, plastic interlockable ring offered stability to the graft during culture, transport, and transplantation. The use of the standardized amniotic membrane, within the amnion ring construct, improves reproducibility of the results and therefore heralds elective surgery. Rapid transplantation of a wrinkle-free graft, using a sutureless, “no touch" technique was achieved and this allowed precise tailoring of the graft to the recipient bed.
This is the first time a standardized, clinical grade protocol has been described for manufacturing limbal epithelial grafts with an efficient surgical technique that prevents postsurgical graft shrinkage and improves corneal integration. The quick, sutureless, and manipulation-free technique ensured transplantation of viable, proliferating limbal epithelial stem cells.
描述一种标准化、无异种的角膜缘上皮干细胞移植物制备方案，以及一种标准化移植的“无接触”手术技术。
比利时安特卫普大学医院。
通过在标准化（胰蛋白酶处理和去除海绵层）羊膜上培养角膜缘上皮干细胞，然后将其拉伸在可互锁羊膜环内，生成缘-羊膜复合移植物。细胞在含有 1%人 AB 血清的 CnT-20 培养基中培养 2 周。在手术准备好的受者角膜上应用纤维蛋白胶，并在一个连续的动作中，将羊膜环内的复合移植物从培养物中转移并置于移植物床上。通过环钻和/或显微手术剪刀在缘水平上切出所需的大小。
轻便、可互锁的塑料环在培养、运输和移植过程中为移植物提供了稳定性。在羊膜环结构内使用标准化羊膜提高了结果的可重复性，因此预示着选择性手术。使用无缝线的“无接触”技术快速移植无皱纹的移植物，并能精确地将移植物裁剪到受者床。
这是首次描述一种标准化、临床级的角膜缘上皮移植物制备方案，以及一种有效的手术技术，可防止术后移植物收缩并改善角膜整合。快速、无缝线和无需操作的技术确保了有活力、增殖的角膜缘上皮干细胞的移植。
