Characterization of Limbal Stem Cell Deficiency by In Vivo Laser Scanning Confocal Microscopy

Deng, Sophie X.; Sejpal, Kunjal; Tang, Qiongyan; Aldave, Anthony J.; Lee, Olivia L; Yu, Fei

doi:10.1001/archophthalmol.2011.378

Cited by 94 publications

(59 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These results may pave the way for successful gene therapy of in vitro expanded LESC for the future transplantation purposes in cases of severe diabetic keratopathy. Because hereditary and acquired LESC deficiency is also associated with the reduction of corneal nerves similar to diabetes 33,34 , the transplantation of healthy LESC-enriched cultures onto the diseased corneas could potentially alleviate the diabetic corneal neuropathy as well.…”

Section: Discussionmentioning

confidence: 99%

Adenoviral Gene Therapy for Diabetic Keratopathy: Effects on Wound Healing and Stem Cell Marker Expression in Human Organ-cultured Corneas and Limbal Epithelial Cells

Kramerov

Saghizadeh

Ljubimov

2016

JoVE

View full text Add to dashboard Cite

The goal of this protocol is to describe molecular alterations in human diabetic corneas and demonstrate how they can be alleviated by adenoviral gene therapy in organ-cultured corneas. The diabetic corneal disease is a complication of diabetes with frequent abnormalities of corneal nerves and epithelial wound healing. We have also documented significantly altered expression of several putative epithelial stem cell markers in human diabetic corneas. To alleviate these changes, adenoviral gene therapy was successfully implemented using the upregulation of c-met proto-oncogene expression and/or the downregulation of proteinases matrix metalloproteinase-10 (MMP-10) and cathepsin F. This therapy accelerated wound healing in diabetic corneas even when only the limbal stem cell compartment was transduced. The best results were obtained with combined treatment. For possible patient transplantation of normalized stem cells, an example is also presented of the optimization of gene transduction in stem cell-enriched cultures using polycationic enhancers. This approach may be useful not only for the selected genes but also for the other mediators of corneal epithelial wound healing and stem cell function.

show abstract

Section: Discussionmentioning

confidence: 99%

Adenoviral Gene Therapy for Diabetic Keratopathy: Effects on Wound Healing and Stem Cell Marker Expression in Human Organ-cultured Corneas and Limbal Epithelial Cells

Kramerov

Saghizadeh

Ljubimov

2016

JoVE

View full text Add to dashboard Cite

show abstract

“…Previous reports have described cellular changes in different stages of LSCD. 15–17 In the current study, we have expanded the investigation of basal cell density in limbal regions. All measurements were obtained in a masked fashion by two independent observers.…”

Section: Discussionmentioning

confidence: 99%

“…15 Briefly, the early stage was characterized by stippling or late fluorescein staining; the intermediate stage was characterized by persistent, late fluorescein staining in a vortex pattern; and the late stage was characterized by the same vortex pattern of staining and a history of cornea epithelial defect or persistent epithelial defect. Representative slit lamp photos are shown in Figure 1.…”

Section: Methodsmentioning

confidence: 99%

“…Two independent, masked observers then proceeded to measure basal cell density in all three images as recommended by the manufacturer and as previously reported. 15 Cell size was calculated by dividing the size of the area by the number of cells within this area. The average cell diameter was then derived from the calculated cell area.…”

Section: Methodsmentioning

confidence: 99%

“…14 On the basis of previous work, corneal basal cell density and subbasal nerve density were identified as potential parameters for the diagnosis of LSCD and for the characterization of the severity of LSCD. 15 In the current study, we investigated whether limbal basal cell density is another parameter in assessing LSCD.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Limbal Basal Cell Density Decreases in Limbal Stem Cell Deficiency

Chan¹,

Chen²,

Rao³

et al. 2015

American Journal of Ophthalmology

View full text Add to dashboard Cite

Purpose To investigate changes in limbal basal epithelial cell density in eyes with limbal stem cell deficiency (LSCD) using in vivo confocal laser scanning microscopy Design retrospective observational comparative study Methods A total of 43 eyes of 30 patients diagnosed with LSCD were included in the study. Ten eyes from normal subjects were included as control. Confocal imaging of the central cornea, and the superior, nasal, inferior and temporal limbus were collected using the Heidelberg Retina Tomograph III Rostock Corneal Module. Basal cell density in all locations was measured by two independent observers. Results The mean basal cell density of the normal group was 9264 ±598 cells/mm2 in the cornea and 7120 ±362 cells/mm2 in the limbus. In the LSCD group, the mean basal cell density in the cornea decreased 31.0% (6389 ±1820 cells/mm2, p<0.001) and in the limbus decreased 23.6% (5440 ±1123 cells/mm2, p<0.001) compared to that in the control. There was a trend of basal cell density decline in more advanced stage of LSCD. The basal cell density declined in the unaffected regions at a similar degree as that in the affected region in sectoral LSCD (p>0.05). The basal cell diameter increased by 24.6% in the cornea (14.7 μm) and by 15.7% in the limbus (15.5 μm) compared to the control. Conclusions Basal cell density in both central cornea and limbus decreases in LSCD. LSCs are affected globally and basal cell density could be used as a parameter to measure LSC function at the early stages of the disease process.

show abstract

K14 + Compound niches are present on the mouse cornea early after birth and expand after debridement wounds

Pajoohesh‐Ganji

Pal‐Ghosh

Tadvalkar

et al. 2015

Developmental Dynamics

View full text Add to dashboard Cite

Background: We previously identified compound niches (CNs) at the limbal:corneal border of the mouse cornea that contain corneal epithelial progenitor cells, express Keratin 8 (K8), and goblet cell mucin Muc5AC. During re‐epithelialization after 2.5 mm epithelial debridement wounds, CNs migrate onto the cornea and expand in number mimicking conjunctivalization. When CNs form during development and whether they express corneal epithelial progenitor cell enriched K14 was not known. Results: To provide insight into corneal epithelial homeostasis, we quantify changes in expression of simple (K8, K18, K19) and stratified squamous epithelial keratins (K5, K12, K14, and K15) during postnatal development and in response to 2.5 mm wounds using quantitative polymerase chain reaction (Q‐PCR), confocal imaging and immunoblots. K14 + CNs are present 7 days after birth. By 21 days, when the eyelids are open, K8, K19, and Muc5AC are also expressed in CNs. By 28 days after wounding, the corneal epithelium shows enhanced mRNA and protein expression for K14 and retains mRNA and protein for corneal epithelial specific K12. Conclusions: The keratin phenotype observed in corneal epithelial cells before eyelid opening is similar to that seen during wound healing. Data show K14 + corneal epithelial progenitor cells expand in number after 2.5 mm wounds. Developmental Dynamics 245:132–143, 2016. © 2015 The Authors. Developmental Dynamics published by Wiley Periodicals, Inc.

show abstract

Characterization of Limbal Stem Cell Deficiency by In Vivo Laser Scanning Confocal Microscopy

Cited by 94 publications

References 24 publications

Adenoviral Gene Therapy for Diabetic Keratopathy: Effects on Wound Healing and Stem Cell Marker Expression in Human Organ-cultured Corneas and Limbal Epithelial Cells

Adenoviral Gene Therapy for Diabetic Keratopathy: Effects on Wound Healing and Stem Cell Marker Expression in Human Organ-cultured Corneas and Limbal Epithelial Cells

Limbal Basal Cell Density Decreases in Limbal Stem Cell Deficiency

K14 + Compound niches are present on the mouse cornea early after birth and expand after debridement wounds

Contact Info

Product

Resources

About