Abstract
Corneal tissue regeneration is of crucial importance for maintaining normal vision. We aimed to isolate and cultivate human corneal stroma-derived mesenchymal stem-like cells (CSMSCs) from the central part of cadaver corneas and study their phenotype, multipotency, role in immunity and wound healing. The isolated cells grew as monolayers in vitro, expressed mesenchymal- and stemness-related surface markers (CD73, CD90, CD105, CD140b), and were negative for hematopoietic markers as determined by flow cytometry. CSMSCs were able to differentiate in vitro into fat, bone and cartilage. Their gene expression profile was closer to bone marrow-derived MSCs (BMMSCs) than to limbal epithelial stem cells (LESC) as determined by high-throughput screening. The immunosuppressive properties of CSMSCs were confirmed by a mixed lymphocyte reaction (MLR), while they could inhibit proliferation of activated immune cells. Treatment of CSMSCs by pro-inflammatory cytokines and toll-like receptor ligands significantly increased the secreted interleukin-6 (IL-6), interleukin-8 (IL-8) and C-X-C motif chemokine 10 (CXCL-10) levels, as well as the cell surface adhesion molecules. CSMSCs were capable of closing a wound in vitro under different stimuli. These cells thus contribute to corneal tissue homeostasis and play an immunomodulatory and regenerative role with possible implications in future cell therapies for treating sight-threatening corneal diseases.
Highlights
MSC- or bone marrow-derived MSC(BMMSC)-like[13]
The in vitro cultured cells fulfilled the ISCT criteria for MSCs20, as they were adherent to the cell culture plastic, could differentiate into three different lineages and expressed the desired MSC markers (CD73, CD90, CD105 and CD140b/PDGFRβ)
The percentage of positive cells for the mannose and D-glucose binding concanavalin A (ConA) was higher in corneal stroma-derived mesenchymal stem-like cells (CSMSCs) cultures, with the concomitant decrease in UEA, PNA, DBA and SBA positivity
Summary
MSC- or bone marrow-derived MSC(BMMSC)-like[13]. Nothing is known about the participation of corneal stroma stem cells in corneal tissue remodelling and immunomodulatory processes related to trauma or infections[17]. We isolated and characterized human central corneal stroma stem cells and compared their genotype to LESCs and BMMSCs, as well as their surface marker phenotype to BMMSCs. In this study, we isolated and characterized human central corneal stroma stem cells and compared their genotype to LESCs and BMMSCs, as well as their surface marker phenotype to BMMSCs Their differentiation potential and the immunological and wound healing properties were tested ex vivo to possibly harvest such cells for future cell, immunosuppressive and wound healing therapy in humans[6]
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