Depth and age-dependent distribution of keratocytes in healthy human corneas

Berlau, J.; Becker, Hans-Henner; Stave, J.; Oriwol, Constanze; Guthoff, Rudolf

doi:10.1016/s0886-3350(01)01227-5

Cited by 74 publications

(48 citation statements)

References 18 publications

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“…2D); therefore a concentration of 12.5 million cells ml −1 was chosen for all further encapsulations. This value falls in the range of reported in vivo keratocyte densities [21].…”

Section: Hydrogel Culturessupporting

confidence: 68%

Keratocyte behavior in three-dimensional photopolymerizable poly(ethylene glycol) hydrogels

et al. 2008

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The goal of this study was to evaluate three-dimensional (3-D) poly(ethylene glycol) (PEG) hydrogels as a culture system for studying corneal keratocytes. Bovine keratocytes were subcultured in DMEM/F-12 containing 10% fetal bovine serum (FBS) through passage 5. Primary keratocytes (P0) and corneal fibroblasts from passages 1 (P1) and 3 (P3) were photoencapsulated at various cell concentrations in PEG hydrogels via brief exposure to light. Additional hydrogels contained adhesive YRGDS and nonadhesive YRDGS peptides. Hydrogel constructs were cultured in DMEM/F-12 with 10% FBS for 2 and 4 weeks. Cell viability was assessed by DNA quantification and vital staining. Biglycan, type I collagen, type III collagen, keratocan and lumican expression were determined by reverse transcriptase-polymerase chain reaction. Deposition of type I collagen, type III collagen and keratan sulfate (KS)-containing matrix components was visualized using confocal microscopy. Keratocytes in a monolayer lost their stellate morphology and keratocan expression, displayed elongated cell bodies, and up-regulated biglycan, type I collagen and type III collagen characteristic of corneal fibroblasts. Encapsulated keratocytes remained viable for 4 weeks with spherical morphologies. Hydrogels supported production of KS, type I collagen and type III collagen matrix components. PEG-based hydrogels can support keratocyte viability and matrix production. 3-D hydrogel culture can stabilize but not restore the keratocyte phenotype. This novel application of PEG hydrogels has potential use in the study of corneal keratocytes in a 3-D environment.

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“…2D); therefore a concentration of 12.5 million cells ml −1 was chosen for all further encapsulations. This value falls in the range of reported in vivo keratocyte densities [21].…”

Section: Hydrogel Culturessupporting

confidence: 68%

Keratocyte behavior in three-dimensional photopolymerizable poly(ethylene glycol) hydrogels

et al. 2008

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show abstract

“…[58][59][60][61][62] Keratoctye density, in addition, has been reported to decrease with increasing age, 60,62,64 however, other authors 57,63 have reported consistency in the corneal keratocyte populations throughout life. In this study, reduction in keratocyte density with age was found to be significant in the posterior stroma only, with a strong trend within the anterior stroma.…”

Section: 39mentioning

confidence: 99%

Quantitative analysis of corneal microstructure in keratoconus utilising in vivo confocal microscopy

Weed

MacEwen

Cox

et al. 2006

Eye

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“…Several ocular surface agerelated changes have been reported in the literature such as [3] reduction in lacrimal secretion and changes on its composition [4]; reduction in functional meibomian glands and changes in lipid secretions [5]; the composition and amount of the tear film changes [6] and the conjunctival development of conjunctivochalasis [7]. Furthermore, the corneal sensitivity is reduced, epithelial and endothelial basement membranes increase its thickness, the number of keratocytes decrease [8] and there is an increased loss of corneal endothelial cells [9]. The incidence and prevalence of ocular diseases as age-related maculopathy, liquefaction of the vitreous, glaucoma, vascular occlusive diseases, cataract and dry eye increase significantly with age [10,11].…”

Section: Introductionmentioning

confidence: 99%