Abstract
PurposeTo reveal the phenotypic differences between human ocular surface stromal cells (hOSSCs) cultured from the corneal, limbal, and scleral compartments.MethodsA comparative analysis of cultured hOSSCs derived from four unrelated donors was conducted by multichromatic flow cytometry for six distinct CD antigens, including the CD73, CD90, CD105, CD166, CD146, and CD34.ResultsThe hOSSCs, as well as the reference cells, displayed phenotypical profiles that were similar in high expression of the hallmark mesenchymal stem cell markers CD73, CD90, and CD105, and also the cancer stem cell marker CD166. Notably, there was considerable variation regarding the expression of CD34, where the highest levels were found in the corneal and scleral compartments. The multi-differentiation potential marker CD146 was also expressed highly variably, ranging from 9% to 89%, but the limbal stromal and endometrial mesenchymal stem cells significantly surpassed their counterparts within the ocular and reference groups, respectively. The use of six markers enabled investigation of 64 possible variants, however, just four variants accounted for almost 90% of all hOSSCs, with the co-expression of CD73, CD90, CD105, and CD166 and a combination of CD146 and CD34. The limbal compartment appeared unique in that it displayed greatest immunophenotype diversity and harbored the highest proportion of the CD146+CD34- pericyte-like forms, but, interestingly, the pericyte-like cells were also found in the avascular cornea.ConclusionOur findings confirm that the hOSSCs exhibit an immunophenotype consistent with that of MSCs, further highlight the phenotypical heterogeneity in stroma from distinct ocular surface compartments, and finally underscore the uniqueness of the limbal region.
Highlights
Continuous maintenance of the cornea is imperative to preserve the normal vision
It has been found that several subtypes of human ocular surface stromal cells display stem cell-like properties, with similar stem celllike features being attributed to stromal cells from avascular central cornea[8] and the sclera.[9]
It has been revealed that human ocular surface stromal cells (hOSSCs) have a phenotype in common with mesenchymal stem cells (MSCs), including the expression of CD29, CD54, CD71, CD90, CD105, CD106, and CD166.[8, 9, 12] The hOSSCs appear to represent a highly functional population, which likely plays an important role in the maintenance of cornea, and in the future may support a new generation of improved therapies for sight-threatening corneal diseases, such as limbal stem cell deficiency or corneal scarring
Summary
Continuous maintenance of the cornea is imperative to preserve the normal vision. It has been shown that the transitional zone between cornea and sclera known as limbus plays an important role,[1, 2] and we have in our previous work highlighted some important aspects of the epithelial stem cells associated with this part of the ocular surface.[3–5] In addition to the limbal epithelial stem cells (LESCs), the limbus harbors a group of stem cell-like stromal cells capable of multilineage differentiation.[6, 7] it has been found that several subtypes of human ocular surface stromal cells (hOSSCs) display stem cell-like properties, with similar stem celllike features being attributed to stromal cells from avascular central cornea[8] and the sclera.[9]. It has been shown that the transitional zone between cornea and sclera known as limbus plays an important role,[1, 2] and we have in our previous work highlighted some important aspects of the epithelial stem cells associated with this part of the ocular surface.[3–5]. It has been revealed that hOSSCs have a phenotype in common with mesenchymal stem cells (MSCs), including the expression of CD29, CD54, CD71, CD90, CD105, CD106, and CD166.[8, 9, 12] The hOSSCs appear to represent a highly functional population, which likely plays an important role in the maintenance of cornea, and in the future may support a new generation of improved therapies for sight-threatening corneal diseases, such as limbal stem cell deficiency or corneal scarring The hOSSCs become mitotic, exhibit limited capacity for self-renewal and transit into myofibroblast phenotype.[10, 11] It has been revealed that hOSSCs have a phenotype in common with mesenchymal stem cells (MSCs), including the expression of CD29, CD54, CD71, CD90, CD105, CD106, and CD166.[8, 9, 12] The hOSSCs appear to represent a highly functional population, which likely plays an important role in the maintenance of cornea, and in the future may support a new generation of improved therapies for sight-threatening corneal diseases, such as limbal stem cell deficiency or corneal scarring
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