April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Human Bone Marrow-Derived Mesenchymal Stem Cells Support the Growth of Limbal Epithelial Progenitor/Stem Cells
Author Affiliations & Notes
  • Sheyla Gonzalez
    Ophthalmology, Jules Stein Eye Institute UCLA, Los Angeles, CA
  • Hua Mei
    Ophthalmology, Jules Stein Eye Institute UCLA, Los Angeles, CA
  • Martin N Nakatsu
    Ophthalmology, Jules Stein Eye Institute UCLA, Los Angeles, CA
  • Elfren Ray Baclagon
    Ophthalmology, Jules Stein Eye Institute UCLA, Los Angeles, CA
  • Sophie Xiaohui Deng
    Ophthalmology, Jules Stein Eye Institute UCLA, Los Angeles, CA
  • Footnotes
    Commercial Relationships Sheyla Gonzalez, UCLA (P); Hua Mei, UCLA (P); Martin Nakatsu, UCLA (P); Elfren Baclagon, None; Sophie Deng, UCLA (P)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 522. doi:https://doi.org/
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      Sheyla Gonzalez, Hua Mei, Martin N Nakatsu, Elfren Ray Baclagon, Sophie Xiaohui Deng; Human Bone Marrow-Derived Mesenchymal Stem Cells Support the Growth of Limbal Epithelial Progenitor/Stem Cells. Invest. Ophthalmol. Vis. Sci. 2014;55(13):522. doi: https://doi.org/.

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

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Abstract

Purpose: To test whether human bone marrow-derived mesenchymal stem cells (BM-MSCs) can serve as feeder cells to expand human limbal epithelial progenitor/stem cells (LSCs).

Methods: Primary human LSCs were isolated and seeded as single cells and cell clusters in the presence of BM-MSCs. LSCs were cultured directly on a BM-MSC monolayer (standard method), in a 3-dimensional (3-D) method, and 3-D fibrin method. Single LSCs cultured on a 3T3-J2 monolayer were used as a control. Cultured LSCs were analyzed for cell morphology, cell growth rate, and expression level of putative stem cell markers.

Results: LSCs did not grow efficiently using the direct or standard method. LSCs in the cell cluster form showed a 2.5-fold higher cell growth rate using the 3-D method than in the control. LSCs cultured by 3-D and 3-D fibrin methods had a similar K14 mRNA expression level and a significantly lower K12 mRNA expression compared to the control (decreased by 34% and 95%, respectively; p<0.05). There was no difference in the percentage of K14+ and K12+ cells among all culture methods. The percentage of p63α bright cells in the 3-D (13.6%) and 3-D fibrin method (11%) was comparable to that in the control (12.8%, p>0.05).

Conclusions: BM-MSCs could support the growth and maintenance of LSCs. The expansion rate was the highest in the novel 3-D culture method.

Keywords: 482 cornea: epithelium • 721 stem cells  
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