Early and late-onset cell migration from peripheral corneal endothelium

PLoS ONE, Год журнала: 2023, Номер 18(5), С. e0285609 - e0285609

Опубликована: Май 10, 2023

In this study we describe peripheral corneal endothelial cell migration in vitro the absence and presence of a ROCK-inhibitor. For study, 21 graft rims, with attached trabecular meshwork (TM), were prepared from Descemet membrane-endothelial sheets by 6.5 mm trepanation. initial proof-of-concept, 7 outer rims cultured thermo-reversible hydrogel matrix for up to 47 days. To assess effect ROCK-inhibitor, 14 paired either or without ROCK-inhibitor 46 At end culture, tissue was retrieved examined viability expression different markers (ZO-1, Na+/K+-ATPase, NCAM, glypican, vimentin). All remained viable displayed single regions (n = 5/21) collective areas 16/21) migration, regardless ROCK-inhibition. Migration started after 4±2 days continued at least 29 The seemed contribute more regular morphology migrating cells. addition, demonstrated phenotypically distinct late-onset but fast-growing population emerging area close limbus. These cells emerged 3 weeks culture appeared less differentiated compared other migration. Immunostaining showed that migrated maintained patterns markers. conclusion, observed 2 morphologically populations first type being triggered broken physical barrier, which disrupted contact inhibition second, showing higher proliferative capacity though appearing differentiated. This subpopulation be mediated stimuli than loss presence. Further exploration differences between these types may assist optimizing regenerative treatment options diseases.

Язык: Английский

---

A single-cell RNA-seq analysis unravels the heterogeneity of primary cultured human corneal endothelial cells

Scientific Reports, Год журнала: 2023, Номер 13(1)

Опубликована: Июнь 8, 2023

Abstract The cornea is a transparent and avascular tissue located in front of the eye. Its inner surface lined by monolayer corneal endothelial cells (CECs), which maintain transparency. CECs remain arrested non-proliferative state damage to these can compromise their function leading opacity. primary culture donor-derived promising cell therapy. It confers potential treat multiple patients from single donor, alleviating global donor shortage. Nevertheless, this approach has limitations preventing its adoption, particularly protocols allow limited expansion there lack clear parameters identify therapy-grade CECs. To address limitation, better understanding molecular changes arising required. Using single-cell RNA sequencing on cultured CECs, we variable transcriptomic fingerprint at level, provide pseudo-temporal reconstruction culture, suggest markers assess quality CEC cultures. This research depicts deep cellular heterogeneity sets basis for further improvement therapies.

Язык: Английский

---

Impact of culture media on primary human corneal endothelial cells derived from old donors

Experimental Eye Research, Год журнала: 2024, Номер 240, С. 109815 - 109815

Опубликована: Фев. 3, 2024

Corneal endothelial dysfunction is a major indication for corneal transplantation. However, global shortage of donor tissues and risks associated with surgeries have prompted exploration alternative options, including tissue-engineered grafts or cell injection therapy. Nonetheless, these approaches require controlled culture primary human cells (HCEnCs). Although HCEnCs established from young donors are generally more proliferative maintain better phenotype, corneas old frequently accessible eye banks due to lower count than the necessary threshold required In this study, we investigated various media evaluate which one most appropriate stimulating proliferation while maintaining morphology function derived (age >65 years). All experiments were performed on paired research-grade corneas, divided conditions under investigation in order minimize inter-donor variability. Cell as well expression specific markers assessed at both mRNA (CD166, SLC4A11, ATP1A1, COL8A1, α-SMA, CD44, COL1A1, CDKN2A, LAP2A LAP2B) protein (ZO-1, Ki67 LAP2) levels. Results obtained showed how Dual Media formulation maintained hexagonal phenotype efficiently Single Medium, but size gradually increased passages. contrast, Medium provided higher rate prolonged vitro expansion acquired an elongated morphology. To summarize, medium preserve functional low passages maintenance same features high remains active area research. The new insights revealed within work become particularly relevant considering that elderly population a) main target therapy, b) represents majority donors. Therefore, proper essential develop novel personalised therapeutic strategies reduce requirement globally.

Язык: Английский

---

Femtosecond Laser Cutting of Human Crystalline Lens Capsule and Decellularization for Corneal Endothelial Bioengineering

Bioengineering, Год журнала: 2024, Номер 11(3), С. 255 - 255

Опубликована: Март 5, 2024

The bioengineering of corneal endothelial grafts consists seeding in vitro cultured cells onto a thin, transparent, biocompatible, and sufficiently robust carrier which can withstand surgical manipulations. This is one the most realistic alternatives to donor corneas, are chronic global shortage. anterior capsule crystalline lens has already been identified as best possible carriers, but its challenging manual preparation limited use. In this study, we describe femtosecond laser cutting process whole lenses order obtain discs 8 mm diameter, similar conventional grafts. Circular marks made on periphery disc indicate orientation. Immersion water for 3 days sufficient completely remove epithelial enable cells, remain viable after 27 culture. Therefore, method provides decellularized ready form tissue engineered

Язык: Английский

---

Tissue Engineering and Ophthalmology

Turkish Journal of Ophthalmology, Год журнала: 2024, Номер unknown, С. 159 - 169

Опубликована: Июнь 28, 2024

Tissue engineering (TE) is a field of science that combines biological, engineering, and medical sciences allows the development disease models, drug gene therapy studies, even cellular or tissue-based treatments developed by methods.The eye an organ easily accessible amenable to applications, paving way for TE in ophthalmology.TE studies are being conducted on wide range topics, including tear film, eyelids, cornea, optic nerve, glaucoma, retinal diseases.With rapid scientific advances field, it seems radically modifying management ocular disorders.

Язык: Английский

---