Sharpe Justin R, Daya Sheraz M, Dimitriadi Maria, Martin Robin, James S Elizabeth
Blond McIndoe Centre, Queen Victoria Hospital, East Grinstead, and Department of Pharmacy and Biomolecular Sciences, University of Brighton, UK.
Tissue Eng. 2007 Jan;13(1):123-32. doi: 10.1089/ten.2006.0108.
In this study a technique for determining donor cell fate following corneal grafting was evaluated. Patients treated for limbal deficiency with allogeneic cultured corneal epithelial cells were studied to determine the fate of the grafted cells. The technique was evaluated initially through the use of donor eyes and then applied to the clinical analysis of 7 patients who had received a cultured corneal epithelial allograft. Cells removed from the cornea and any retrieved tissue were analyzed via polymerase chain reaction (PCR) genotyping to determine the origin of the cells populating the patients' healed cornea. A mixture of genotypes was detected in a cornea retrieved from a patient following a fully penetrating keratoplasty who had received a mixture of allogeneic tissue. Donor cells were no longer detected on the corneal surface of all 7 cases beyond 28 weeks postgraft. At these later time points, only patient genotype could be detected. These results demonstrate that PCR genotyping can be used to determine the origin of cells populating the surface of the cornea following the grafting of cultured allogeneic cells and demonstrates that transplanted cultured limbal epithelial cells do not persist on the surface of the host cornea for more than 28 weeks.
在本研究中,对一种用于确定角膜移植后供体细胞命运的技术进行了评估。对接受同种异体培养角膜上皮细胞治疗角膜缘缺陷的患者进行研究,以确定移植细胞的命运。该技术最初通过使用供体眼进行评估,然后应用于7例接受培养角膜上皮同种异体移植患者的临床分析。从角膜移除的细胞和任何回收的组织通过聚合酶链反应(PCR)基因分型进行分析,以确定构成患者愈合角膜的细胞来源。在一名接受了同种异体组织混合物的穿透性角膜移植术后患者的回收角膜中检测到了混合基因型。在移植后28周以上,所有7例患者的角膜表面均未再检测到供体细胞。在这些较晚的时间点,只能检测到患者基因型。这些结果表明,PCR基因分型可用于确定培养的同种异体细胞移植后构成角膜表面的细胞来源,并表明移植的培养角膜缘上皮细胞在宿主角膜表面持续时间不超过28周。