Current Perspectives of Limbal-Derived Stem Cells and its Application in Ocular Surface Regeneration and Limbal Stem Cell Transplantation

Singh, Vivek; Tiwari, Anil; Kethiri, Abhinav Reddy; Sangwan, Veena

doi:10.1002/sctm.20-0408

Cited by 29 publications

(16 citation statements)

References 81 publications

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“…Therefore, the graft rejection-related problems and the limited availability of suitable donors represent major hurdles for successful allogeneic LSC transplantations. To address this problem, alternative autologous tissue sources such as hair follicle and dental pulp stem cells, oral mucosal epithelia, and mesenchymal stem cells have been investigated for their potential for corneal regeneration upon transplantation [ 5 , 6 , 10 ]. However, despite eliminating the need for immunosuppression, incomplete differentiation towards the corneal epithelial phenotype may pose a problem for using alternative autologous tissue grafts [ 10 ].…”

Section: Introductionmentioning

confidence: 99%

Corneal epithelial differentiation of human pluripotent stem cells generates ABCB5+ and ∆Np63α+ cells with limbal cell characteristics and high wound healing capacity

et al. 2021

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Background Differentiation of functional limbal stem cells (LSCs) from human pluripotent stem cells (hPSCs) is an important objective which can provide novel treatment solutions for patients suffering from limbal stem cell deficiency (LSCD). Yet, further characterization is needed to better evaluate their immunogenicity and regenerative potential before clinical applications. Methods Human PSCs were differentiated towards corneal fate and cryopreserved using a clinically applicable protocol. Resulting hPSC-LSC populations were examined at days 10–11 and 24–25 during differentiation as well as at passage 1 post-thaw. Expression of cornea-associated markers including PAX6, ABCG2, ∆Np63α, CK15, CK14, CK12 and ABCB5 as well as human leukocyte antigens (HLAs) was analyzed using immunofluorescence and flow cytometry. Wound healing properties of the post-thaw hPSC-LSCs were assessed via calcium imaging and scratch assay. Human and porcine tissue-derived cultured LSCs were used as controls for marker expression analysis and scratch assays at passage 1. Results The day 24–25 and post-thaw hPSC-LSCs displayed a similar marker profile with the tissue-derived LSCs, showing abundant expression of PAX6, ∆Np63α, CK15, CK14 and ABCB5 and low expression of ABCG2. In contrast, day 10–11 hPSC-LSCs had lower expression of ABCB5 and ∆Np63α, but high expression of ABCG2. A small portion of the day 10–11 cells coexpressed ABCG2 and ABCB5. The expression of class I HLAs increased during hPSC-LSCs differentiation and was uniform in post-thaw hPSC-LSCs, however the intensity was lower in comparison to tissue-derived LSCs. The calcium imaging revealed that the post-thaw hPSC-LSCs generated a robust response towards epithelial wound healing signaling mediator ATP. Further, scratch assay revealed that post-thaw hPSC-LSCs had higher wound healing capacity in comparison to tissue-derived LSCs. Conclusions Clinically relevant LSC-like cells can be efficiently differentiated from hPSCs. The post-thaw hPSC-LSCs possess functional potency in calcium responses towards injury associated signals and in wound closure. The developmental trajectory observed during hPSC-LSC differentiation, giving rise to ABCG2+ population and further to ABCB5+ and ∆Np63α+ cells with limbal characteristics, indicates hPSC-derived cells can be utilized as a valuable cell source for the treatment of patients afflicted corneal blindness due to LSCD. Graphical Abstract

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Section: Introductionmentioning

confidence: 99%

Corneal epithelial differentiation of human pluripotent stem cells generates ABCB5+ and ∆Np63α+ cells with limbal cell characteristics and high wound healing capacity

et al. 2021

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“…Examples of success in this approach include limbal and macular stem cell implants used in ophthalmic procedures. 77 , 78 Within the vocal fold, mesenchymal stem cells are most commonly used to restore lamina propria as they retain the capacity for protein secretion, can remodel matrix protein and are believed to reduce the hosts fibrotic response via a paracrine effect involving growth factors and exosome nanoparticles. 12 , 79 , 80 Current challenges with this approach include enabling cell survival following implantation with most reports lacking a description of the long‐term fate of implanted cells.…”

Section: Resultsmentioning

confidence: 99%

The life‐cycle and restoration of the human vocal fold

Hamilton

2022

Laryngoscope Investig Oto

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Objective: To better understand the challenges of designing therapies to treat damaged vocal fold lamina propria, it is essential to understand the biophysical and pathophysiological mechanisms involved in vocal fold development, maintenance, injury, and aging. This review critically analyses these points to try and direct future efforts and new strategies toward science-based solutions. Data Sources & Review Methods: MEDLINE, Ovid Embase, and Wed of Science databases were used to identify relevant literature. A scoping review was performed following the preferred reporting items for systematic reviews and meta-analyses extension for scoping reviews checklist. Results: The layered arrangement of the vocal fold, develops during early childhood and is maintained during adulthood unless injury occurs. The stellate cells of the macular flava are likely to be important in this process. The capacity for vocal fold regeneration and growth is lost during adulthood and repair results in the deposition of fibrous tissue from resident fibroblasts. With advancing age, viscoelastic tissue declines, possibly due to cell senescence. Strategies aimed at replacing fibrous tissue within the vocal folds must either stimulate resident cells or implant new cells to secrete healthy extracellular protein. Injection of basic fibroblast growth factor is the most widely reported therapy that aims to achieve this. Conclusions: The pathways involved in vocal fold development, maintenance and aging are incompletely understood. Improved understanding has the potential to identify new treatment targets that could potentially overcome loss of vocal fold vibratory tissue.

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“…However, CK3 is a marker to identify corneal epithelial cells specifically and CK19 is used to identify conjunctival epithelial cells exclusively. [ 11 12 ] The absence of limbal stem cell markers could be because the samples were obtained from the central 7.75 mm of the cornea (keratoplasty samples) without encroaching on to the limbal region. However, the presence of progenitor cells is a noteworthy finding as it implies that there is some expression of stem cell activity, which may have the capability to restore limbal stem cell functionality even in the sequelae stage of the disease.…”

Section: Discussionmentioning

confidence: 99%

Clinical, histological and immunohistochemistry characteristics of cornea in the sequelae stage of chronic vernal keratoconjunctivitis

Jaffet

Singh²,

Chaurasia³

et al. 2021

Indian Journal of Ophthalmology

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Purpose: To report the clinical outcomes and histopathological and immunohistochemistry (IHC) features in eyes with the sequelae stage of vernal keratoconjunctivitis (VKC). Methods: Investigative study of corneal samples obtained following surgical intervention for vision restoration in four eyes of three patients with VKC. Patient 1 (an 11-year-old boy) had deep anterior lamellar keratoplasty in both eyes, Patient 2 (a 24-year-old male) underwent superficial keratectomy followed by penetrating keratoplasty, and Patient 3 (a 22-year-old male) underwent penetrating keratoplasty. The corneal samples retrieved after surgical intervention were assessed for histology features and immunohistochemistry (IHC) studies. Results: The grafts were clear till the follow-up of 2–18 months. Histopathology of all four corneal samples showed epithelial hyperplasia, absent Bowman layer, thick hyalinized stromal lamellae, vascularization, and chronic inflammatory cells such as lymphocytes and plasma cells. IHC showed strong expression of CK 3 in both eyes of Patient 1 and no expression in Patients 2 and 3. The marker for limbal stem cells, ABCG2, was absent in all four samples; however, p63α was expressed strongly in Patients 2 and 3, moderately in the right eye of Patient 1, and marginally expressed in the left eye of Patient 1. Conclusion: The eyes in the sequelae stage of VKC (having corneal scarring and 360° hypertrophied limbus) can be managed favorably with keratoplasty and amniotic membrane transplantation without allogenic/cadaveric stem cell transplantation. The expression of transient progenitor cells in the scarred corneas of VKC patients in the sequelae stage suggests that the limbal stem cell dysfunction is more likely partial and self-renewal of limbal stem cells is a plausibility in these eyes.

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Current Perspectives of Limbal-Derived Stem Cells and its Application in Ocular Surface Regeneration and Limbal Stem Cell Transplantation

Cited by 29 publications

References 81 publications

Corneal epithelial differentiation of human pluripotent stem cells generates ABCB5+ and ∆Np63α+ cells with limbal cell characteristics and high wound healing capacity

Corneal epithelial differentiation of human pluripotent stem cells generates ABCB5+ and ∆Np63α+ cells with limbal cell characteristics and high wound healing capacity

The life‐cycle and restoration of the human vocal fold

Clinical, histological and immunohistochemistry characteristics of cornea in the sequelae stage of chronic vernal keratoconjunctivitis

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