Isolation and Culture of Corneal Stromal Stem Cells

Nagymihály, Richárd; Moe, Morten C.; Petrovski, Goran

doi:10.1007/978-1-0716-0599-8_1

Cited by 7 publications

(7 citation statements)

References 17 publications

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“…Isolation of corneal stromal fibroblasts from adult rats was done using a corneal explant protocol (Nagymihaly & Moe, 2020 ). Briefly the adult male rats were sacrificed, and the eyeballs were dissected out in a petri dish in PBS.…”

Section: Methodsmentioning

confidence: 99%

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Chitosan-coated bovine serum albumin nanoparticles for topical tetrandrine delivery in glaucoma: in vitro and in vivo assessment

et al. 2022

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Glaucoma is one of the leading causes of blindness. Therapies available suffer from several drawbacks including low bioavailability, repeated administration and poor patient compliance with adverse effects thereafter. In this study, bovine serum albumin nanoparticles (BSA-NPs) coated with chitosan(CS) were developed for the topical delivery of tetrandrine (TET) for glaucoma management. Optimized nanoparticles were prepared by desolvation. pH, BSA, CS and cross-linking agent concentrations effects on BSA-NPs colloidal properties were investigated. CS-BSA-NPs with particle size 237.9 nm and zeta potential 24 mV was selected for further evaluation. EE% exceeded 95% with sustained release profile. In vitro mucoadhesion was evaluated based on changes in viscosity and zeta potential upon incubation with mucin. Ex vivo transcorneal permeation was significantly enhanced for CS coated formulation. In vitro cell culture studies on corneal stromal fibroblasts revealed NPs biocompatibility with enhanced cellular uptake and improved antioxidant and anti-proliferative properties for the CS-coated formulation. Moreover, BSA-NPs were nonirritant as shown by HET-CAM test. Also, bioavailability in rabbit aqueous humor showed 2-fold increase for CS-TET-BSA-NPs compared to TET with a sustained reduction in intraocular pressure in a rabbit glaucoma model. Overall, results suggest CS-BSA-NPs as a promising platform for topical ocular TET delivery in the management of glaucoma.

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Section: Methodsmentioning

confidence: 99%

“…The media were refreshed every 3–4 days. Once spindle adherent cells from tissue explants reached 80-90% confluency, they were trypsinized using 0.25% trypsin EDTA (Lonza, Basel, Switzerland) and expanded in larger T75 cm 2 flasks (Nagymihaly & Moe, 2020 ).…”

Section: Methodsmentioning

confidence: 99%

Chitosan-coated bovine serum albumin nanoparticles for topical tetrandrine delivery in glaucoma: in vitro and in vivo assessment

et al. 2022

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“…[ 51,52 ] The cells in the scaffolds cannot overgrow, therefore, they will not transdifferentiate and lose their MSC phenotype. [ 53 ] While the presence of Col I and Col V appears to support the hypothesis that the cells are producing their own ECM and the fibrillogenesis process is ongoing. [ 34,35,40–42 ] No immediate adverse reaction to the implants could be observed at day 14; however, it must be noted, that in some cases, the corneal flap could not close properly around the edges of the excisions, and therefore, no apparent healing process could be observed.…”

Section: Discussionmentioning

confidence: 87%

Intracorneal Implantation of 3D Bioprinted Scaffolds Containing Mesenchymal Stromal Cells Using Femtosecond‐Laser‐Assisted Intrastromal Keratoplasty

Boix‐Lemonche

Nagymihály

Niemi

et al. 2023

Macromolecular Bioscience

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Injury of the cornea is a complex biological process. Regeneration of the corneal stroma can be facilitated by the presence of mesenchymal stromal cells (MSCs) and application of tissue equivalents. A new tissue‐engineering strategy for corneal stroma regeneration is presented using cellularized 3D bioprinted hydrogel constructs implanted into organ cultured porcine corneas using femtosecond laser‐assisted intrastromal keratoplasty. The ex vivo cultured, MSC‐loaded 3D bioprinted structures remain intact, support cell survival, and contain de novo synthesized extracellular matrix components and migrating cells throughout the observation period. At day 14 postimplantation, the cellularized tissue equivalents contain few or no cells, as demonstrated by optical coherence tomography imaging and immunofluorescent staining. This study successfully combines a laboratory‐based method with modern, patient‐care practice to produce a cell‐laden tissue equivalent for corneal implantation. Optimal bioink composition and cellularization of tissue equivalents are essential in fine‐tuning a method to promote the current technique as a future treatment modality.

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“…The transplantation of healthy keratocyte progenitor cells into keratoconus corneas would provide a novel treatment modality that may slow the progression of keratoconus [ 21 ]. Moreover, the corneal stromal stem cells, a rare cell population resident in the peripheral cornea and limbus, can be isolated by specific surface markers from limbal stromal tissues [ 22 – 25 ]. Du et al injected the human corneal stromal stem cells into mice corneas and did not observe elicit immune rejection over an extended period of time, suggesting an opportunity to develop cell-based therapies for corneal stromal diseases [ 26 ].…”

Section: Strategies For Corneal Stromal Regenerationmentioning

confidence: 99%

New dawn for keratoconus treatment: potential strategies for corneal stromal regeneration

Dou,

Liu,

Shi

et al. 2023

Stem Cell Res Ther

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Keratoconus is a progressive, ectatic and blinding disorder of the cornea, characterized by thinning of corneal stroma. As a highly prevalent among adolescents, keratoconus has been a leading indication for corneal transplantation worldwide. However, the severe shortage of donor corneas is a global issue, and the traditional corneal transplantation surgeries may superinduce multiple complications, necessitating efforts to develop more effective strategies for keratoconus treatment. In this review, we summarized several strategies to promote corneal stromal regeneration or improve corneal stromal thickness, including cell-based therapies, biosynthetic alternatives for inducing corneal regeneration, minimally invasive intrastromal implantation and bioengineered tissues for implantation. These strategies provided more accessible but safer alternatives from various perspectives for keratoconus treatment, paving the way for arresting the keratoconus progression in its earlier stage. For the treatments of corneal ectatic diseases beyond keratoconus, these approaches will provide important references and widen the therapy options in a donor tissue-independent manner.

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Isolation and Culture of Corneal Stromal Stem Cells

Cited by 7 publications

References 17 publications

Chitosan-coated bovine serum albumin nanoparticles for topical tetrandrine delivery in glaucoma: in vitro and in vivo assessment

Chitosan-coated bovine serum albumin nanoparticles for topical tetrandrine delivery in glaucoma: in vitro and in vivo assessment

Intracorneal Implantation of 3D Bioprinted Scaffolds Containing Mesenchymal Stromal Cells Using Femtosecond‐Laser‐Assisted Intrastromal Keratoplasty

New dawn for keratoconus treatment: potential strategies for corneal stromal regeneration

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