September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Rate of Proliferation in Corneal Limbal versus Bone Marrow Mesenchymal Stem Cells
Author Affiliations & Notes
  • Ali R Djalilian
    Ophthalmology, Univ of Illinois at Chicago, Chicago, Illinois, United States
  • Samaneh Ghassemi
    Ophthalmology, Univ of Illinois at Chicago, Chicago, Illinois, United States
  • Gaurav Agnihotri
    Ophthalmology, Univ of Illinois at Chicago, Chicago, Illinois, United States
  • Judy Hamouie
    Ophthalmology, Univ of Illinois at Chicago, Chicago, Illinois, United States
  • Asha Tadepalli
    Ophthalmology, Univ of Illinois at Chicago, Chicago, Illinois, United States
  • Ilham Putra
    Ophthalmology, Univ of Illinois at Chicago, Chicago, Illinois, United States
  • Xiang Shen
    Ophthalmology, Univ of Illinois at Chicago, Chicago, Illinois, United States
  • Peiman Hematti
    Hematology, University of Wisconsin, Madison, Wisconsin, United States
  • Medi Eslani
    Ophthalmology, Univ of Illinois at Chicago, Chicago, Illinois, United States
  • Footnotes
    Commercial Relationships   Ali Djalilian, None; Samaneh Ghassemi, None; Gaurav Agnihotri, None; Judy Hamouie, None; Asha Tadepalli, None; Ilham Putra, None; Xiang Shen, None; Peiman Hematti, None; Medi Eslani, 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; Research funding and corneal tissue from Eversight; and unrestricted grant to the department from RPB.
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 4923. doi:
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    • Get Citation

      Ali R Djalilian, Samaneh Ghassemi, Gaurav Agnihotri, Judy Hamouie, Asha Tadepalli, Ilham Putra, Xiang Shen, Peiman Hematti, Medi Eslani; Rate of Proliferation in Corneal Limbal versus Bone Marrow Mesenchymal Stem Cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):4923.

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

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Abstract

Purpose : Mesenchymal stem cells (MSC) are a promising source of cells for cell based therapies in part based on their regenerative and immunomodulatory properties. MSCs have been isolated from many tissues such as bone marrow, fat, dental pulp, cornea, etc. In this study, we compared the rate of proliferation between Corneal Limbal derived MSCs (CL-MSC) and bone marrow derived MSCs (BM-MSC).

Methods : MSCs were isolated from healthy human bone marrow donors and from human corneal limbus from research eyebank corneas (generously provided by Eversight). They were plated in 175 cm2 flasks at a seeding density of 2000 cells/cm2 in MEM-alpha with 10% fetal bovine serum. The cells were serially passaged at 80% confluency and doubling time was calculated. Cell proliferation at mid log proliferation phase were determined using the Click-iT EdU kit (Thermo-fisher) by fluorescent-activated cell sorting (BD LSR Fortessa). All experiments were repeated with at least five different donors.

Results : The average doubling time of CL-MSC was 14.6 ± 1.8, 56.7 ± 16.2, 44.0 ± 1.4, 38.6 ± 10.8 and 36.9 ± 3.1 hours in passage 2, 3, 4, 5 and 6, respectively. For BM-MSC the doubling time was 64.6 ± 15.5, 83.7 ±12.3, 76.2 ± 17.0, 72.8 ± 11.0 and 75.3 ± 13.5 hours in passage 2, 3, 4, 5, and 6, respectively, which was significantly longer than CL-MSCs (p value for each comparison < 0.01). Incubation with 10 mM EdU for 24 hours resulted in 100.0 ± 5.1, 62.2 ± 18.1, 76.5 ± 14.4, 83.8 ± 12.7 and 76.2 ± 17.4 percent of EdU positive cells in passage 2 to 6 of CL-MSC, respectively. On the other hand, in BM-MSC 38.4 ± 9.7, 31.8 ± 12.1, 34.8 ± 11.5, 27.4 ± 16.3 and 38.5 ± 14.9 percent were EdU positive after 24 hours incubation (p value for each comparison < 0.01).

Conclusions : These results demonstrate CL-MSCs have a faster proliferation rate compared to BM-MSCs and can be expanded in culture in greater numbers. Given the availability of eyebank corneal tissue and ease of isolation and rate of proliferation, CL-MSCs can be a suitable alternative for cell based therapy especially for ocular surface diseases.

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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