Human Limbal Mesenchymal Stem Cells Express
            <i>ABCB5</i>
            and Can Grow on Amniotic Membrane

Shaharuddin, Bakiah; Osei-Bempong, Charles; Ahmad, Sajjad; Rooney, Paul; Ali, Simi; Meeson, Annette

doi:10.2217/rme-2016-0009

Cited by 18 publications

(19 citation statements)

References 56 publications

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“…Modeling RPE functional changes, for example basolateral functions, that take place in pathological states like in cases of macular degeneration are more preferable in the form of 3D culture and cell spheroids 15 . The limbal stroma of the eye is one of the promising sources of MSCs 16 . Bioprinting approaches in ophthalmology are used primarily in reconstruction of the anterior segment with the production of artificial cornea and lens that are unique and patient-specific in shape and design 17 .…”

mentioning

confidence: 99%

Cell spheroid fusion: beyond liquid drops model

Kosheleva

Efremov

Shavkuta

et al. 2020

Sci Rep

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Biological self-assembly is crucial in the processes of development, tissue regeneration, and maturation of bioprinted tissue-engineered constructions. The cell aggregates-spheroids-have become widely used model objects in the study of this phenomenon. existing approaches describe the fusion of cell aggregates by analogy with the coalescence of liquid droplets and ignore the complex structural properties of spheroids. Here, we analyzed the fusion process in connection with structure and mechanical properties of the spheroids from human somatic cells of different phenotypes: mesenchymal stem cells from the limbal eye stroma and epithelial cells from retinal pigment epithelium. A nanoindentation protocol was applied for the mechanical measurements. We found a discrepancy with the liquid drop fusion model: the fusion was faster for spheroids from epithelial cells with lower apparent surface tension than for mesenchymal spheroids with higher surface tension. this discrepancy might be caused by biophysical processes such as extracellular matrix remodeling in the case of mesenchymal spheroids and different modes of cell migration. The obtained results will contribute to the development of more realistic models for spheroid fusion that would further provide a helpful tool for constructing cell aggregates with required properties both for fundamental studies and tissue reparation. Modern approaches to the rapidly evolving fields of regenerative medicine and tissue engineering are closely associated with the development and formation of tissue-engineered constructions, where cellular components play a crucial role 1-3. Monolayer cell culture is the most widely used approach to the growing and studying of cells in vitro. Nevertheless, 2D culture conditions cause cell flattening and remodeling of the cell's internal structure, which can eventually affect the gene expression 4. On the other hand, 3D cell culture better reflects the in vivo microenvironment both morphologically and physiologically. The extra dimension which 3D cell cultures have, compared to monolayers, helps to establish intercellular junctions, to reorganize the cytoskeleton, to polarize and to differentiate in conditions similar to native tissue conditions 5. Multicellular spheroids obtained under nonadhesive conditions represent one possible 3D cell culture system. There is a great deal of unexplored potential in spheroid-based research, as tissue engineering using spheroids is a relatively new field 6-8. Three-dimensional bioprinting of scaffold-based and scaffold-free tissue-engineered constructions is widely used for tissue substitution and modeling of organs-on-chips 9-12. Cell spheroids with prefabricated intercellular junctions and extracellular matrix provide a new promising type of bioinks suitable for processing by an

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mentioning

confidence: 99%

Cell spheroid fusion: beyond liquid drops model

Kosheleva

Efremov

Shavkuta

et al. 2020

Sci Rep

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“…However, our results showed stromal staining of ABCB5 in the limbus, and some LESCs were ABCB5 + /p63α – . A recent study reported that human limbal mesenchymal stem cells also expressed ABCB5 [ 40 ]. In addition, previous studies found that ABCB5 was expressed in skin progenitor cells and melanoma stem cells, and functioned as a regulator of cellular differentiation [ 41 , 42 ].…”

Section: Discussionmentioning

confidence: 99%

“…These data raise questions: whether ABCB5 only marks LESCs in the limbus and whether ABCB5 is a definitive marker of LESCs. Interestingly, two widely proposed LESC markers, p63α and ABCG2, were also expressed in human limbal mesenchymal stem cells [ 40 ], and ABCG2 was also expressed in Langerhans cells rather than LESCs in the limbus [ 43 ]. Thus, we believe that all of these proposed LESC markers are not definitive cell markers only for LESCs.…”

Section: Discussionmentioning

confidence: 99%

Poly(ethylene glycol)-modified silk fibroin membrane as a carrier for limbal epithelial stem cell transplantation in a rabbit LSCD model

Yang

et al. 2017

Stem Cell Res Ther

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BackgroundLimbal epithelial stem cells (LESCs) play important roles in corneal epithelial homeostasis and regeneration, and damage to the limbus will lead to limbal stem cell deficiency (LSCD), with conjunctivalization and even visual impairment. Cultured LESCs have been used for ocular surface reconstruction, and silk fibroin (SF) membranes have shown potential as a substrate for LESC cultivation. Both culture methods and the carriers of LESCs affect outcomes following LESC transplantation.MethodsRabbit LESCs were cultured from tissue explant, single cell-suspension, and cell cluster culture methods. Ratios of p63α and/or ABCB5-positive LESCs, differentiated corneal epithelial cells (CK12 staining), and corneal tight junction formation (Claudin-1 staining) were examined to choose the most applicable LESC cultures. SF membranes were prepared and modified by 400-Da poly(ethylene glycol) (PEG). The characteristics of stem cells and normal corneal differentiation of LESCs cultured on PEG-modified SF membranes were further examined by immunofluorescence staining and flow cytometric analysis. LESCs cultured on PEG-modified SF membranes (LESC/SF grafts) and PEG-modified SF membranes (SF grafts) were transplanted onto rabbit corneas with total LSCD. New blood vessels, corneal epithelial defects, and cornea clarity were examined after transplantation. Furthermore, corneal epithelial thickness, stromal thickness, and the percentage area of CK12-positive corneal epithelium were quantified 4 months after transplantation.ResultsTissue explant and single cell-suspension cultures harvested more p63α and/or ABCB5-positive LESCs, generated more CK12-positive corneal epithelial cells, and formed more corneal tight junctions than cell cluster cultures. Prepared PEG-modified SF membranes were transparent, flexible, and sturdy enough for surgical manipulation. LESCs cultured on PEG-modified SF membranes maintained characteristics of stem cells and normal corneal differentiation. LESC/SF grafts inhibited new blood vessels and rescued corneal epithelial defects in the rabbit total LSCD model. In addition, LESC/SF grafts repopulated the limbus and increased corneal epithelial thickness, stromal thickness, and the area percentage of CK12-positive corneal epithelium.ConclusionsLESCs from tissue explant and single cell-suspension cultures were more applicable corneal epithelial cells for ocular surface reconstruction. LESC/SF grafts repaired corneal epithelial defects and reversed LSCD, and PEG-modified SF membranes were suitable to be a carrier for LESC transplantation.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-017-0707-y) contains supplementary material, which is available to authorized users.

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“…It has also been suggested that MSCs produce growth factors that can support the growth of residual corneal epithelial cells and LESCs [ 95 ]. Recent researches by Shaharuddin et al found that cultured limbal MSCs expressed the common putative limbal stem cell markers, which demonstrated limbal-derived MSC-exhibited plasticity [ 96 ].…”

Section: Cell-based Therapymentioning

confidence: 99%

Emerging Therapeutic Strategies for Limbal Stem Cell Deficiency

Dong

Han

Lavker

2018

Journal of Ophthalmology

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Identification and characterization of the limbal epithelial stem cells (LESCs) has proven to be a major accomplishment in anterior ocular surface biology. These cells have been shown to be a subpopulation of limbal epithelial basal cells, which serve as the progenitor population of the corneal epithelium. LESCs have been demonstrated to play an important role in maintaining corneal epithelium homeostasis. Many ocular surface diseases, including intrinsic (e.g., Sjogren's syndrome) or extrinsic (e.g., alkali or thermal burns) insults, which impair LESCs, can lead to limbal stem cell deficiency (LSCD). LSCD is characterized by an overgrowth of conjunctival-derived epithelial cells, corneal neovascularization, and chronic inflammation, eventually leading to blindness. Treatment of LSCD has been challenging, especially in bilateral total LSCD. Recently, advances in LESC research have led to novel therapeutic approaches for treating LSCD, such as transplantation of the cultured limbal epithelium. These novel therapeutic approaches have demonstrated efficacy for ocular surface reconstruction and restoration of vision in patients with LSCD. However, they all have their own limitations. Here, we describe the current status of LSCD treatment and discuss the advantages and disadvantages of the available therapeutic modalities.

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Human Limbal Mesenchymal Stem Cells Express ABCB5 and Can Grow on Amniotic Membrane

Abstract: Limbal-derived MSC exhibited plasticity, could maintain limbal markers expression and demonstrated viable growth on amniotic membrane.

Cited by 18 publications

References 56 publications

Cell spheroid fusion: beyond liquid drops model

Cell spheroid fusion: beyond liquid drops model

Poly(ethylene glycol)-modified silk fibroin membrane as a carrier for limbal epithelial stem cell transplantation in a rabbit LSCD model

Emerging Therapeutic Strategies for Limbal Stem Cell Deficiency

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