Interactions between stem cells and their microenvironment are critical for regulation maintenance of cell function. To elucidate the molecular interactions within human limbal epithelial stem/progenitor (LEPC) niche, which is essential maintaining corneal transparency vision, we performed a comprehensive expression analysis adhesion molecules (CAMs) using custom-made quantitative real-time polymerase chain reaction (qRT-PCR) arrays laser capture-microdissected LEPC clusters, comprising...

Allogenic transplants of the cornea are prone to rejection, especially in repetitive transplantation and scarred or highly vascularized recipient sites. Patients with these ailments would particularly benefit from possibility use non-immunogenic decellularized tissue scaffolds for transplantation, which may be repopulated by host cells situ vitro. So, aim this study was develop a fast efficient decellularization method creating human corneal extracellular matrix scaffold suitable...

Background: Bone marrow mesenchymal cells have been identified as a source of pluripotent stem with varying degrees plasticity in humans. However, there are few reports on rat-derived cells, which could be good models for the research purpose. We describe here simple method establishing rat bone stromal by principle adhesion and document their phenotype along differentiation potential to other lineages. Materials Methods: Rat were isolated three methods: direct plastic adherence, ficoll...

Abstract Understanding transcription factor (TF) regulation of limbal epithelial stem/progenitor cells (LEPCs) may aid in using non-ocular to regenerate the corneal surface. This study aimed identify and characterize TF genes expressed specifically LEPCs isolated from human donor eyes by laser capture microdissection. Using a profiling approach, preferential expression was found for SoxE SoxF genes, particularly Sox9, which showed predominantly cytoplasmic localization basal nuclear...

Optimization of culture conditions for human limbal epithelial stem/progenitor cells (LEPC) that incorporate the in vivo cell-matrix interactions are essential to enhance LEPC ex vivo-expansion and transplantation efficiency. Here, we investigate efficacy laminin (LN) isoforms preferentially expressed niche as matrices tissue engineering. Analyses expression patterns LN chains provided evidence enrichment LN-α2, -α3, -α5, -β1, -β2, -β3, -γ1, -γ2 -γ3 basement membrane, with LN-α5 representing...

Limbal melanocytes (LMel) represent essential components of the corneal epithelial stem cell niche and are known to protect limbal stem/progenitor cells (LEPCs) from UV damage by transfer melanosomes. Here, we explored additional functional roles for LMel in homeostasis, immune regulation angiostasis. Human corneoscleral tissues were morphologically analyzed normal, inflammatory wound healing conditions. The effects on LEPCs direct indirect co-culture models using electron microscopy,...

Abstract The myocardium is unable to regenerate itself after infarct, resulting in scarring and thinning of the heart wall. Our objective was develop a patch buttress bypass scarred area, while allowing regeneration by incorporated cardiac stem/progenitor cells (CPCs). Polycaprolactone (PCL) fabricated as both sheets solvent casting, fibrous meshes electrospinning, potential patches, determine role topology proliferation phenotypic changes CPCs. Thiophene‐conjugated carbon nanotubes (T‐CNTs)...

Abstract PAX3, a transcription factor essential for neural crest development and melanocyte progenitors, is expressed in various melanocytic tissues. However, its role ocular surface tissues remains poorly understood. This study investigated the expression patterns of PAX3 limbal stem cell niche, specifically epithelial progenitor cells (LEPC), melanocytes (LM), mesenchymal (LMSC). Additionally, was studied conjunctival/limbal melanoma specimens. Immunohistochemical analysis revealed...

Paired box 6 (PAX6), a nuclear transcription factor, determines the fate of limbal epithelial progenitor cells (LEPC) and maintains cell identity. However, expression PAX6 in niche cells, primarily mesenchymal stromal (LMSC), melanocytes is scarce not entirely clear. To distinctly assess fresh organ-cultured human corneoscleral tissues were stained immunohistochemically. Furthermore, cultured was investigated. Immunostaining revealed presence PAX6-negative which positive for vimentin...

Limbal epithelial progenitor cells (LEPC) rely on their niche environment for proper functionality and self-renewal. While extracellular vesicles (EV), specifically small EVs (sEV), have been proposed to support LEPC homeostasis, data sEV derived from limbal like mesenchymal stromal (LMSC) remain limited, there are no studies sEVs melanocytes (LM). In this study, we isolated conditioned media of LMSC LM using a combination tangential flow filtration size exclusion chromatography...

Abstract Limbal melanocytes, located in the basal epithelial layer of corneoscleral limbus, represent essential components corneal stem cell niche, but, due to difficulties their isolation and cultivation, biological roles potential for cell-based tissue engineering approaches have not been comprehensively studied. Here, we established a protocol efficient cultivation pure populations human limbal which could be expanded at high yield by using recombinant laminin (LN)-511-E8 as culture...

Despite great progress in the identification of mesenchymal stem cells (MSCs) from bone marrow (BM), our knowledge their vivo cellular identity remains limited. We report here that expressing transcription factor Ebf2 adult BM display characteristics MSCs. The Ebf2(+) are highly clonal and physiologically quiescent. In lineage-tracing experiments, single cell clone transplantations, vitro differentiation assays revealed self-renewal multilineage capacity. Gene expression analysis freshly...

The transplantation of ex vivo expanded limbal epithelial progenitor cells (LEPCs) on amniotic membrane or fibrin gel is an established therapeutic strategy to regenerate the damaged corneal surface in patients with stem cell deficiency (LSCD), but long-term success rate restricted. A scaffold niche-specific structure and extracellular matrix (ECM) composition might have advantage improve clinical outcomes, particular for severe damage complete loss niche tissue structure. Therefore, we...

The fate decision of limbal epithelial progenitor cells (LEPC) at the human corneal limbus is determined by surrounding microenvironment with niche (LNC) as one its essential components. Research on freshly isolated LNC which mainly include mesenchymal stromal (LMSC) and melanocytes (LM) has been hampered a lack efficient protocols to isolate purify these cells. We devised protocol for rapid retrieval pure LMSC, LM LEPC populations collagenase digestion tissue subsequent...

The porcine ocular surface is used as a model of the human surface; however, detailed characterization has not been documented. This due, in part, to scarcity antibodies produced specifically against cell types or structures. We performed histological and immunohistochemical investigation on frozen formalin-fixed, paraffin-embedded tissue from domestic pigs using panel 41 different related epithelial progenitor/differentiation phenotypes, extracellular matrix associated molecules, various...

Organ culture storage techniques for corneoscleral limbal (CSL) tissue have improved the quality of corneas transplantation and allow longer times. Cultured has been used stem cell to treat deficiency (LSCD) as well research purposes assess homeostasis mechanisms in niche. However, effects organ conditions on niche components are less described. Therefore, this study, morphological immunohistochemical characteristics organ-cultured investigated compared fresh tissues by means light electron...

Abstract Limbal melanocytes (LM) are located in the basal epithelial layer of corneoscleral limbus and interact with adjacent limbal progenitor cells. The exploration their biological role maintenance stem cell niche has been limited by difficulty LM isolation cultivation. Here, we report on a facile protocol for efficient enrichment pure populations human LMs fluorescence-activated sorting (FACS) using antibodies raised against surface marker CD117 (c-Kit). enriched retain self-renewal...

Interactions between limbal epithelial progenitor cells (LEPC) and surrounding niche cells, which include mesenchymal stromal (LMSC) melanocytes (LM), are essential for the maintenance of stem cell required a transparent corneal surface. P-cadherin (P-cad) is critical adhesion molecule at various niches; however, conflicting observations were reported on presence P-cad in region. To explore this issue, we assessed location phenotype P-cad+ by confocal microscopy human corneoscleral tissue....