Optimization of human corneal endothelial cells for culture: the removal of corneal stromal fibroblast contamination using magnetic cell separation. | LitMetric
The culture of human corneal endothelial cells (CECs) is critical for the development of suitable graft alternative on biodegradable material, specifically for endothelial keratoplasty, which can potentially alleviate the global shortage of transplant-grade donor corneas available. However, the propagation of slow proliferative CECs in vitro can be hindered by rapid growing stromal corneal fibroblasts (CSFs) that may be coisolated in some cases. The purpose of this study was to evaluate a strategy using magnetic cell separation (MACS) technique to deplete the contaminating CSFs from CEC cultures using antifibroblast magnetic microbeads. Separated "labeled" and "flow-through" cell fractions were collected separately, cultured, and morphologically assessed. Cells from the "flow-through" fraction displayed compact polygonal morphology and expressed Na(+)/K(+)ATPase indicative of corneal endothelial cells, whilst cells from the "labeled" fraction were mostly elongated and fibroblastic. A separation efficacy of 96.88% was observed. Hence, MACS technique can be useful in the depletion of contaminating CSFs from within a culture of CECs.
A male patient in his 20s, suffering from a persistent, infection-related corneal endothelial plaque (EP) was urgently referred to our tertiary medical centre for therapeutic penetrating keratoplasty (TPK). Over the preceding month, he had been undergoing treatment with both topical and oral antifungal medications due to clinical suspicion of fungal keratitis. At our centre, an endothelial scraping was performed using a reverse Sinskey hook to obtain samples for microbiology and revealed septate branching fungal hyphae.
Purpose: To compare corneal endothelial loss in manual small incision cataract surgery (MSICS) following three different techniques of nucleus delivery (Viscoexpression, Irrigating Vectis, Phacosandwich).
Setting: Mahatma Gandhi Medical College and Research Institute, Puducherry, India.
Design: Prospective, single blinded, randomized controlled trial.
Introduction: The purpose of this study was to compare the growth factor concentrations in undiluted autologous platelet-rich plasma (APRP) and autologous serum (AS) eye drops.
Methods: This was a single-center, prospective trial conducted in a tertiary university hospital in Bangkok, Thailand. Ninety-six patients with moderate-to-severe dry eye disease, who were randomly assigned to receive either 100% APRP or 100% AS, were enrolled in the study.
Purpose: To assess the safety and the efficacy of the "Sub-400 corneal cross-linking (CXL) protocol" for progressive keratoconus (KC) in ultrathin corneas.
Methods: The study included thirty four patients with progressive KC, who underwent CXL using the "Sub-400" protocol due to intraoperative thinnest corneal pachymetry ranging from 295 to 398 μm after epithelial removal. After the epithelium was removed, the following ultraviolet A irradiation was applied at a fluence of 3 mW/cm and the duration was adjusted based on the specific corneal stromal thickness.
Aim: To observe the clinical outcomes of 30-gauge (G) needle-guided 10-0 prolene suture purse-string pupilloplasty for atonic pupil management.
Methods: Eight patients with atonic pupils who underwent suture purse-string pupilloplasty were retrospectively analyzed. Preoperative data and at least 6mo of postoperative data were collected from all the patients.
