Timolol induces necroptosis, apoptosis and senescence concentration-dependently in rabbit Limbal stem cells in vitro

Currently, a helium-neon laser is widely used in the treatment of degenerative, inflammatory and vascular diseases of the eye. It was found that the interaction of this radiation with different tissues, as a result of complex photochemical processes, is manifested by anti-inflammatory, desensitizing, resolving effects. Also, there is a stimulating effect on the processes of reparation and trophism. However, these observations relate to the use of laser for external irradiation, when the light goes through the air and the sclera or through the cornea. Research of the effect of endo-laser direct exposure of the fiber in the cavity of the eye, where light acts directly on epithelial or limbal cells, wasn’t carry out due to objective reasons (difficulty of conducting such research directly to patients). There is currently no data on the effect of helium-neon laser on the limbal area of the eye-ball, enriched endogenous stem and progenitor cells. They help repair damaged tissues in the eye (anterior and posterior segment) at various injuries. Cell culture, grown in vitro, are free from the influences of body systems, attract researchers as a unique model to study the behavior of cells in normal but also in response to external or internal factors. Currently, scientists have learned how to obtain and difficult to cultivate a culture of cells, in-cluding tissues of the human eye. They are genetically homogeneous population of cells growing in constant conditions. The viability and morphology of cells and their ultra-structure and various molecular biological cha-racteristics can be studied in cell culture, and to find the optimal conditions of laser irradiation, does not damage the cells. In the present work the authors studied the effect of low-energy helium-neon laser irradiation (632 nm) on limbal stem cells. Conducted in vitro studies have shown that the use of low-energy helium-neon laser irrad-iation (632 nm) has a positive effect on the monolayer culture of limbal stem cells. Absence of changes in cell phenotype and high proliferative activity indicate a stimulating effect of the investigated radiation on stem and progenitor cells, leading to the result of the activation of recovery and reduction of pathological changes at the cellular and organ and tissue levels.

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In vitro cultivation of porcine limbal stem cells

Italian Journal of Animal Science ◽

10.4081/ijas.2009.s3.125 ◽

2009 ◽

Vol 8 (sup3) ◽

pp. 125-127

Author(s):

Maja Popović ◽

Mirna Tominac ◽

Ksenija Vlahović ◽

Dubravko Kezić ◽

Marcela Šperanda ◽

...

Keyword(s):

Stem Cells ◽

In Vitro Cultivation ◽

Limbal Stem Cells

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Corneal stromal mesenchymal stem cells: reconstructing a bioactive cornea and repairing the corneal limbus and stromal microenvironment

International Journal of Ophthalmology ◽

10.18240/ijo.2021.03.19 ◽

2021 ◽

Vol 14 (3) ◽

pp. 448-455

Author(s):

Xian-Ning Liu ◽

◽

Yun Chen ◽

Yao Wang ◽

◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Anterior Part ◽

Corneal Stroma ◽

Limbal Stem Cells ◽

Differentiation Potential ◽

Self Renewal ◽

Stromal Microenvironment ◽

Application Prospects

Corneal stroma-derived mesenchymal stem cells (CS-MSCs) are mainly distributed in the anterior part of the corneal stroma near the corneal limbal stem cells (LSCs). CS-MSCs are stem cells with self-renewal and multidirectional differentiation potential. A large amount of data confirmed that CS-MSCs can be induced to differentiate into functional keratocytes in vitro, which is the motive force for maintaining corneal transparency and producing a normal corneal stroma. CS-MSCs are also an important component of the limbal microenvironment. Furthermore, they are of great significance in the reconstruction of ocular surface tissue and tissue engineering for active biocornea construction. In this paper, the localization and biological characteristics of CS-MSCs, the use of CS-MSCs to reconstruct a tissue-engineered active biocornea, and the repair of the limbal and matrix microenvironment by CS-MSCs are reviewed, and their application prospects are discussed.

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Characterization of the secretory profile and exosomes of limbal stem cells in the canine species

PLoS ONE ◽

10.1371/journal.pone.0244327 ◽

2020 ◽

Vol 15 (12) ◽

pp. e0244327

Author(s):

Antonio J. Villatoro ◽

Cristina Alcoholado ◽

María del Carmen Martín-Astorga ◽

Gustavo Rico ◽

Viviana Fernández ◽

...

Keyword(s):

Stem Cells ◽

Ocular Surface ◽

Inhibitory Effect ◽

Proteomic Profile ◽

Limbal Stem Cells ◽

High Concentrations ◽

First Time

Limbal stem cells (LSCs) are a quiescent cell population responsible for the renewal of the corneal epithelium. Their deficiency is responsible for the conjunctivization of the cornea that is seen in different ocular pathologies, both in humans and in the canine species. The canine species represents an interesting preclinical animal model in ocular surface pathologies. However, the role of LSCs in physiological and pathological conditions in canine species is not well understood. Our objective was to characterize for the first time the soluble factors and the proteomic profile of the secretome and exosomes of canine LSCs (cLSCs). In addition, given the important role that fibroblasts play in the repair of the ocular surface, we evaluated the influence of the secretome and exosomes of cLSCs on their proliferation in vitro. Our results demonstrated a secretory profile of cLSCs with high concentrations of MCP-1, IL-8, VEGF-A, and IL-10, as well as significant production of exosomes. Regarding the proteomic profile, 646 total proteins in the secretome and 356 in exosomes were involved in different biological processes. Functionally, the cLSC secretome showed an inhibitory effect on the proliferation of fibroblasts in vitro, which the exosomes did not. These results open the door to new studies on the possible use of the cLSC secretome or some of its components to treat certain pathologies of the ocular surface in canine species.

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In Vitro Generation of Pancreatic Endocrine Cells from Human Adult Fibroblast-Like Limbal Stem Cells

Cell Transplantation ◽

10.3727/096368911x580635 ◽

2012 ◽

Vol 21 (1) ◽

pp. 73-90 ◽

Cited By ~ 10

Author(s):

Angela Criscimanna ◽

Giovanni Zito ◽

Annalisa Taddeo ◽

Pierina Richiusa ◽

Maria Pitrone ◽

...

Keyword(s):

Stem Cells ◽

Endocrine Cells ◽

Limbal Stem Cells ◽

Human Adult ◽

Pancreatic Endocrine Cells

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An important role for adenine, cholera toxin, hydrocortisone and triiodothyronine in the proliferation, self-renewal and differentiation of limbal stem cells in vitro

Experimental Eye Research ◽

10.1016/j.exer.2016.09.008 ◽

2016 ◽

Vol 152 ◽

pp. 113-122 ◽

Cited By ~ 10

Author(s):

Min Yu ◽

Sanja Bojic ◽

Gustavo S. Figueiredo ◽

Paul Rooney ◽

Julian de Havilland ◽

...

Keyword(s):

Stem Cells ◽

Cholera Toxin ◽

Limbal Stem Cells ◽

Self Renewal

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Paracrine activity of adipose derived stem cells on limbal epithelial stem cells

10.21203/rs.3.rs-551083/v1 ◽

2021 ◽

Author(s):

Bartosz Sikora ◽

Aleksandra Skubis-Sikora ◽

Agnieszka Prusek ◽

Joanna Gola

Keyword(s):

Stem Cells ◽

Culture Cell ◽

Elisa Test ◽

Inflammatory Factors ◽

Epithelial Stem Cells ◽

Limbal Stem Cells ◽

Paracrine Effect ◽

Expression Of Genes ◽

Limbal Epithelial Stem Cells

Abstract Limbal stem cells deficiency (LSCD) is an eye disease caused by the loss of stem cells in the corneal limbus as a succession of an injury due physical, biological, or chemical agents. Current therapies of LSCD are focused on the transplantation of donor corneas or tissue equivalents produced from autologous limbal stem cells. Every year there are waiting millions of people for the cornea transplantation all over the world and the list of waiting patients is growing due to the relatively low number of cornea donors. On the other hand, the transplantation of tissue or cells into the recipient’s body is associated with the higher risk of possible side effects. The possibility of the application of an indirect treatment using the properties of the paracrine activity of stem cells, would be beneficial for the patients with transplant failures. This study was to evaluate the paracrine effect of mesenchymal stem cells derived from adipose tissue (ADSC) on the viability of limbal epithelial stem cells (LESC). The paracrine effect was assessed by treating LESC with conditioned medium collected from ADSC culture. Cell viability, cytotoxicity, apoptosis and proliferation were evaluated using in vitro assays in standard conditions and induced inflammation. After the exposure to the examined conditions, the expression of genes related to pro- and anti- inflammatory factors was evaluated and compared to the secretion of selected cytokines by ELISA test. Moreover, the changes in LESC phenotype were assessed using of phenotype microarrays. Our findings suggest that paracrine activity of ADSC on LESC promotes its proliferation and mitigates the adverse impact of induced inflammation.

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Time-course single-cell RNA sequencing reveals transcriptional dynamics and heterogeneity of limbal stem cells derived from human pluripotent stem cells

Cell & Bioscience ◽

10.1186/s13578-021-00541-4 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Changbin Sun ◽

Hailun Wang ◽

Qiwang Ma ◽

Chao Chen ◽

Jianhui Yue ◽

...

Keyword(s):

Stem Cells ◽

Single Cell ◽

Cell Fate ◽

Time Course ◽

Embryonic Stem ◽

Developmental Trajectory ◽

Rna Seq ◽

Limbal Stem Cells ◽

Developmental Origin

Abstract Background Human pluripotent stem cell-derived limbal stem cells (hPSC-derived LSCs) provide a promising cell source for corneal transplants and ocular surface reconstruction. Although recent efforts in the identification of LSC markers have increased our understanding of the biology of LSCs, much more remains to be characterized in the developmental origin, cell fate determination, and identity of human LSCs. The lack of knowledge hindered the establishment of efficient differentiation protocols for generating hPSC-derived LSCs and held back their clinical application. Results Here, we performed a time-course single-cell RNA-seq to investigate transcriptional heterogeneity and expression changes of LSCs derived from human embryonic stem cells (hESCs). Based on current protocol, expression heterogeneity of reported LSC markers were identified in subpopulations of differentiated cells. EMT has been shown to occur during differentiation process, which could possibly result in generation of untargeted cells. Pseudotime trajectory analysis revealed transcriptional changes and signatures of commitment of hESCs-derived LSCs and their progeny—the transit amplifying cells. Conclusion Single-cell RNA-seq revealed time-course expression changes and significant transcriptional heterogeneity during hESC-derived LSC differentiation in vitro. Our results demonstrated candidate developmental trajectory and several new candidate markers for LSCs, which could facilitate elucidating the identity and developmental origin of human LSCs in vivo.

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Paracrine activity of adipose derived stem cells on limbal epithelial stem cells

Scientific Reports ◽

10.1038/s41598-021-99435-1 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Bartosz Sikora ◽

Aleksandra Skubis-Sikora ◽

Agnieszka Prusek ◽

Joanna Gola

Keyword(s):

Stem Cells ◽

Culture Cell ◽

Elisa Test ◽

In Vitro Assays ◽

Inflammatory Factors ◽

Epithelial Stem Cells ◽

Limbal Stem Cells ◽

Paracrine Effect ◽

Limbal Epithelial Stem Cells

AbstractLimbal stem cells deficiency (LSCD) is an eye disease caused by the loss of stem cells in the corneal limbus as a succession of an injury due physical, biological, or chemical agents. Current therapies of LSCD are focused on the transplantation of donor corneas or tissue equivalents produced from autologous limbal stem cells. Every year there are waiting millions of patients for the cornea transplantation all over the world and the list is growing due to the relatively low number of cornea donors. On the other hand, the transplantation of tissue or cells into the recipient’s body is associated with the higher risk of possible side effects. The possibility of the application of an indirect treatment using the properties of the paracrine activity of stem cells, would be beneficial for the patients with transplant failures. This study was to evaluate the paracrine effect of mesenchymal stem cells derived from adipose tissue (ADSC) on the viability of limbal epithelial stem cells (LESC). The paracrine effect was assessed by treating LESC with conditioned medium collected from ADSC culture. Cell viability, cytotoxicity, apoptosis and proliferation were evaluated using in vitro assays in standard conditions and induced inflammation. After the exposure to the examined conditions, the expression of genes related to pro- and anti- inflammatory factors was evaluated and compared to the secretion of selected cytokines by ELISA test. Moreover, the changes in LESC phenotype were assessed using of phenotype microarrays. Our findings suggest that paracrine activity of ADSC on LESC promotes its proliferation and has a potential role in mitigation of the adverse impact of inflammation induced by lipopolysaccharide.

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