July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Comparative genetic analysis of human mesenchymal stem cells isolated from corneal stroma and bone marrow
Author Affiliations & Notes
  • Steffi Matthyssen
    Ophthalmology, Visual Optics and Visual Rehabilitation, Translational Neurosciences, University of Antwerp, Wilrijk, Belgium
  • Veerle Van Gerwen
    Ophthalmology, Visual Optics and Visual Rehabilitation, Translational Neurosciences, University of Antwerp, Wilrijk, Belgium
  • Carina Koppen
    Ophthalmology, Visual Optics and Visual Rehabilitation, Translational Neurosciences, University of Antwerp, Wilrijk, Belgium
    Ophthalmology, Antwerp University Hospital, Edegem, Belgium
  • Nadia Zakaria
    Ophthalmology, Visual Optics and Visual Rehabilitation, Translational Neurosciences, University of Antwerp, Wilrijk, Belgium
    Ophthalmology, Antwerp University Hospital, Edegem, Belgium
  • Footnotes
    Commercial Relationships   Steffi Matthyssen, None; Veerle Van Gerwen, None; Carina Koppen, None; Nadia Zakaria, Novartis Institute for Biomedical Research (E)
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 2276. doi:
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    • Get Citation

      Steffi Matthyssen, Veerle Van Gerwen, Carina Koppen, Nadia Zakaria; Comparative genetic analysis of human mesenchymal stem cells isolated from corneal stroma and bone marrow. Invest. Ophthalmol. Vis. Sci. 2018;59(9):2276.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Corneal stroma-derived mesenchymal stem cells (CS-MSC) have not yet been directly compared on a genetic level with bone marrow MSC (BM-MSC), with or without the inclusion of xenofree conditions. This results in inconclusiveness with regards to the MSC-like properties of corneal stromal cells. The potential of corneal-derived MSCs in tissue engineering applications for Ophthalmology is yet to be explored. We hypothesize that CS-MSC show comparable expression of key MSC genes to BM-MSC, in accordance to findings on a phenotypical and functional level, and that xenofree culture does not alter genetic expression patterns in CS-MSC.

Methods : Human CS-MSC were obtained from six donors and expanded in both standard conditions (basal medium with the addition of 10% Fetal Bovine Serum – FBS) and xenofree conditions using 10% Human Platelet Lysate (HPL) as a replacement for FBS. At passage 4, the cells were harvested and RNA isolation was followed by cDNA extraction and qPCR for 85 key genes for human MSC. Human BM-MSCs at passage 6 (provided by Veneto Eye Bank) were used as control. Phenotypic characterization using flow cytometry and trilineage differentiation was performed to confirm the MSC-like properties of corneal stroma-derived cells as required by the International Society for Cellular Therapy (ISCT).

Results : Out of 85 genes tested, 71 showed no differential expression between bone marrow-derived and corneal stroma-derived MSCs in normal or xenofree conditions. Of the 14 genes where expression differed between BM- and CS-MSCs, 12 were not expressed in BM. Genes with variable expression include: ABCB1, IL10, PROM1, CSF3, IL1B, TNF, KDR, SOX2, BMP7, HNF1A, FGF10 and WNT3A. Flow cytometry showed that corneal stroma-derived MSCs, like BM-SCs, met the ISCT criteria and were positive for CD73, CD90, CD105, CD13, CD29, CD44, CD166 and negative for CD11b, CD 14, CD19, CD34, CD45, CD79a, HLA-DR and were able to differentiate into osteocytes, chondrocytes and adipocytes regardless of the cultures being xenofree or not.

Conclusions : Despite portraying an identical phenotype as described by the ISCT criteria, MSCs derived from the corneal stroma show a different genetic profile to those that are derived from bone marrow. Further research is warranted in order to determine if CS-MSCs would be superior to BM-MSCs for corneal stromal tissue engineering.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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