Human Umbilical Cord-Derived Mesenchymal Stem Cells Promote Corneal Epithelial Repair In Vitro

Corneal injuries are among the leading causes of blindness and vision impairment. Trauma, infectious keratitis, thermal and chemical (acids and alkali burn) injuries may lead to irreversible corneal scarring, neovascularization, conjunctivalization, and limbal stem cell deficiency. Bilateral blindness constitutes 12% of total global blindness and corneal transplantation remains a stand-alone treatment modality for the majority of end-stage corneal diseases. However, global shortage of donor corneas, the potential risk of graft rejection, and severe side effects arising from long-term use of immunosuppressive medications, demands alternative therapeutic approaches. Umbilical cord-derived mesenchymal stem cells can be isolated in large numbers using a relatively less invasive procedure. However, their role in injury induced corneal repair is largely unexplored. Here, we isolated, cultured and characterized mesenchymal stem cells from human umbilical cord, and studied the expression of mesenchymal (CD73, CD90, CD105, and CD34), ocular surface and epithelial (PAX6, WNT7A, and CK-8/18) lineage markers through immunofluorescence. The cultured human limbal and corneal epithelial cells were used as controls. Scratch assay was used to study the corneal epithelial repair potential of umbilical cord-derived mesenchymal stem cells, in vitro. The in vitro cultured umbilical cord-derived mesenchymal stem cells were plastic adherent, showed trilineage differentiation and expressed: mesenchymal markers CD90, CD105, CD73; epithelial marker CK-8/18, and ocular lineage developmental markers PAX6 and WNT-7A. Our findings suggest that umbilical cord-derived mesenchymal stem cells promote repair of the injured corneal epithelium by stimulating the proliferation of corneal epithelial cells, in vitro. They may serve as a potential non-ocular source of stem cells for treating injury induced bilateral corneal diseases.

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Human umbilical cord mesenchymal stem cells improve generation of spheres in human corneal epithelial cells and promotes cell migration

Frontiers in Bioengineering and Biotechnology ◽

10.3389/conf.fbioe.2016.02.00019 ◽

2016 ◽

Vol 4 ◽

Author(s):

Azmi Siti ◽

Kamal Nur Zawani ◽

Hoe Ngai ◽

Mohamed Rafeezul ◽

Jie Tan ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cell Migration ◽

Epithelial Cells ◽

Umbilical Cord ◽

Human Umbilical Cord ◽

Corneal Epithelial Cells ◽

Corneal Epithelial ◽

Human Corneal Epithelial Cells

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Human umbilical cord-mesenchymal stem cells: a promising strategy for corneal epithelial regeneration

Regenerative Medicine ◽

10.2217/rme-2019-0103 ◽

2020 ◽

Vol 15 (3) ◽

pp. 1381-1397 ◽

Cited By ~ 2

Author(s):

Siti Maisura Azmi ◽

Mohamed Salih ◽

Samar Abdelrazeg ◽

Fatin Fazrina Roslan ◽

Rafeezul Mohamed ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Epithelial Cells ◽

Umbilical Cord ◽

Cellular Migration ◽

Human Umbilical Cord ◽

Corneal Epithelial Cells ◽

Corneal Epithelial ◽

Human Telomerase ◽

Epithelial Growth

Aim: As a strategy to improve the outcome of ex vivo cultivated corneal epithelial transplantation, the role of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) is investigated in promoting corneal epithelial growth and functions. Materials & methods: Human telomerase-immortalized corneal epithelial cells were characterized and its functions evaluated by scratch migration assay, cellular senescence, HLA expression and spheres formation with hUC-MSC. Results: Expression of corneal epithelial markers was influenced by the duration and method of co-culture. Indirect co-culture improved cellular migration and delayed senescence when treated after 3 and 5 days. hUC-MSC downregulated expression of HLA Class I and II in IFN-γ-stimulated human telomerase-immortalized corneal epithelial cells. Conclusion: hUC-MSC promote corneal epithelial growth and functions after treatment with hUC-MSC.

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In vitro potential of human mesenchymal stem cells for corneal epithelial regeneration

Regenerative Medicine ◽

10.2217/rme-2019-0067 ◽

2020 ◽

Vol 15 (3) ◽

pp. 1409-1426 ◽

Cited By ~ 2

Author(s):

Núria Nieto-Nicolau ◽

Beatriz Martín-Antonio ◽

Claudia Müller-Sánchez ◽

Ricardo P Casaroli-Marano

Keyword(s):

Stem Cells ◽

Wound Healing ◽

Mesenchymal Stem Cells ◽

Epithelial Cells ◽

Electric Resistance ◽

Corneal Epithelial Cells ◽

Corneal Epithelial ◽

Epithelial Regeneration ◽

Human Corneal Epithelial Cells

Aim: To determine the potential of mesenchymal stem cells (MSC) for corneal epithelial regeneration in vitro. Materials & methods: Bone marrow MSC (BM-MSC) and adipose tissue MSC were analyzed for corneal epithelial and mesenchymal markers, using limbal stem cells and corneal cells as controls. MSC with better potential were cultured with specific mediums for epithelial induction. Transepithelial electric resistance and wound healing assay with human corneal epithelial cells were performed. Results: BM-MSC showed better potential, increased corneal markers, and higher transepithelial electric resistance values when induced with limbal epithelial culture medium. Induced BM-MSC promoted better wound healing of human corneal epithelial cells by paracrine secretion. Conclusion: BM-MSC has potential for corneal epithelial induction in a protocol compatible with human application.

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Umbilical Cord Mesenchymal Stem Cells-derived Exosomes Deliver Mir-21 to Accelerate Corneal Epithelial Wound Healing Through PTEN/PI3K/Akt pathway

10.21203/rs.3.rs-571150/v1 ◽

2021 ◽

Author(s):

Xuran Li ◽

Xiaolong Liu ◽

Yanyan Zhang ◽

Zhiyu Liu ◽

Xinyue Li ◽

...

Keyword(s):

Stem Cells ◽

Wound Healing ◽

Mesenchymal Stem Cells ◽

Umbilical Cord ◽

Wound Repair ◽

Recipient Cell ◽

Human Umbilical Cord ◽

Corneal Epithelial ◽

Epithelial Wound Healing

Abstract BackgroundRapid restoration of corneal epithelium integrity after injury is particularly important for preserving corneal transparency and vision. Mesenchymal stem cells (MSCs) can be taken into account as the promising regenerative therapeutics for improvement of wound healing processes based on the variety of the effective components. The extracellular vesicles form MSCs, especially exosomes, has been considered as important paracrine mediators though transferring microRNAs into recipient cell. This study investigated the mechanism of human umbilical cord MSC-derived exosomes (HUMSC-exosomes) on corneal epithelial wound healing.MethodsExosomes extracted from HUMSCs were identified by transmission electron microscopy, nanoparticle tracking analysis, and Western blot. Corneal fluorescein staining and histological staining were evaluated in a corneal mechanical wound model. Changes in HCECs proliferation after HUMSC-exosomes or miR-21 mimic treatment were evaluated by CCK-8 and EdU assays, while migration was assessed by in vitro scratch wound assay. Full-length transcriptome sequencing was performed to identify the differentially expressed genes associated with HUMSC-exosomes treatment, followed by validation via real-time PCR and Western blot.ResultsThe exosomes derived from HUMSCs can significantly promote corneal epithelial cells proliferation, migration in vitro and accelerate corneal epithelial wound healing in vivo. Similar effects were obtained after miR-21 transfection, while the beneficial effects of HUMSC-exosomes were partially negated by miR-21 knockdown. Results also show that the benefits are associated with decreased PTEN level and activated the PI3K/Akt signaling pathway in HCECs.ConclusionsHUMSC-exosomes could accelerate the recovery of corneal epithelial wounds though restraining PTEN by transferring miR-21, and may represent a promising novel therapeutic agent for corneal wound repair.

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