September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Comparative Effects of Corneal Limbal Versus Bone Marrow Derived Mesenchymal Stem Cell Secretome on Innate Immune Response of Corneal Epithelium
Author Affiliations & Notes
  • Judy Hamouie
    Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago, Illinois, United States
  • Medi Eslani
    Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago, Illinois, United States
  • Samaneh Ghassemi
    Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago, Illinois, United States
  • Gaurav Agnihotri
    Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago, Illinois, United States
  • Asha Tadepalli
    Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago, Illinois, United States
  • Elham Ghahari
    Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago, Illinois, United States
  • Peiman Hematti
    Division of Hematology/Oncology, Department of Medicine, , University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin, United States
  • Ali R Djalilian
    Ophthalmology and Visual Sciences, University of Illinois at Chicago, Chicago, Illinois, United States
  • Footnotes
    Commercial Relationships   Judy Hamouie, None; Medi Eslani, None; Samaneh Ghassemi, None; Gaurav Agnihotri, None; Asha Tadepalli, None; Elham Ghahari, None; Peiman Hematti, None; Ali Djalilian, None
  • Footnotes
    Support  Supported in part by Clinical Scientist Development Program Award K12EY021475 (ME), R01 EY024349-01A1 (ARD) and Core grant EY01792 from NEI/NIH; MR130543 (ARD) from DoD; and unrestricted grant to the department from RPB.
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 1436. doi:
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      Judy Hamouie, Medi Eslani, Samaneh Ghassemi, Gaurav Agnihotri, Asha Tadepalli, Elham Ghahari, Peiman Hematti, Ali R Djalilian; Comparative Effects of Corneal Limbal Versus Bone Marrow Derived Mesenchymal Stem Cell Secretome on Innate Immune Response of Corneal Epithelium. Invest. Ophthalmol. Vis. Sci. 2016;57(12):1436.

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      © 2017 Association for Research in Vision and Ophthalmology.

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Abstract

Purpose : The anti-inflammatory effects of mesenchymal stem cells (MSCs) have been recognized as one of their major modes of action and the basis for their application in a number of human clinical trials. We used our in vitro model of innate immunity (based on toll-like receptor-3 [TLR3] activation) to compare the immunomodulatory effects of Corneal Limbal derived MSC Secretome (CLMSCS) versus Bone Marrow derived MSC Secretome (BMSCS).

Methods : Passage 4 to 6 CLMSC or BMSC were grown to confluency using MEM-alpha + 10% fetal bovine serum. The secretome was collected after 48 hours. It was normalized based on total protein content measurement. Telomerase-immortalized human corneal epithelial cells (HCECs) were stimulated with 1 µg/ml Poly:IC (PIC), a synthetic ligand for TLR3, for 30 minutes, washed with PBS and incubated in either CLMSCS/BMSCS or MEM-alpha for six hours, followed by RNA extraction. After reverse transcription, the mRNA expression was analyzed with ΔΔCt method by real-time qPCR. All experiments were repeated with at least five different donors.

Results : TLR3 activation of HCECs resulted in 2.7 ± 1.1, 4.8 ± 0.6, 5.4 ± 1.9, 27.6 ± 7.6, and 1717.5 ± 28.6 fold increase in IL6, CCL2, CCL3, CCL5, and CXCL10 mRNA expression, respectively (P < 0.001). BMSCS significantly decreased mRNA expression of stimulated HCECs to 1.3 ± 0.8, 2.6 ± 0.8, 3.7 ± 0.8, 4.4 ± 2.1, and 242.5 ± 13.2 fold after TLR3 activation, respectively (P value for each comparison < 0.01). Likewise, CLMSCS significantly reduced these values to 1.5 ± 0.9, 2.9 ± 0.7, 3.2 ± 0.6, 4.1 ± 1.1, and 155.1 ± 21.1 fold, respectively (P < 0.01). There were no significant differences between CLMSCS and BMSCS except for CXCL10, in which CLMSCS resulted in a more inhibition in mRNA expression (P <0.001).

Conclusions : Secretome from CLMSC and BMSC are able to modulate the innate inflammatory response to TLR3 activation in HCECs. They can be used potentially as an anti-inflammatory agent for ocular surface disease.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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