June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Characterization of Mesenchymal Stem Cells vs. Trabecular Meshwork Cells
Author Affiliations & Notes
  • Eric Snider
    Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA
    Biomedical Engineering, Emory University, Atlanta, GA
  • Christopher Pride
    Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA
  • Akash Patil
    Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA
  • W Daniel Stamer
    Biomedical Engineering, Duke University, Durham, NC
  • C Ross Ethier
    Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA
    Biomedical Engineering, Emory University, Atlanta, GA
  • Footnotes
    Commercial Relationships Eric Snider, None; Christopher Pride, None; Akash Patil, None; W Stamer, None; C Ethier, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 2256. doi:
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    • Get Citation

      Eric Snider, Christopher Pride, Akash Patil, W Daniel Stamer, C Ross Ethier; Characterization of Mesenchymal Stem Cells vs. Trabecular Meshwork Cells. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):2256.

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

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Abstract
 
Purpose
 

Due to reduced trabecular meshwork (TM) cellularity in glaucoma, meshwork repair with suitably differentiated stem cells has potential. This requires characterization methods to detect differences between TM and stem cells. The gold standard for identifying TM cells is dexamethasone (dex) induction of myocilin (MYOC); however, to assess differentiation, additional assessments of function and genetic profile are desirable. Here we evaluate multiple techniques to assess differences between mesenchymal stem cells (MSCs) and TM cells.

 
Methods
 

Human adipose-derived MSCs (Lonza; n=3 strains) and primary human TM cells (Stamer lab; n = 4 strains) were used. Three approaches assessed MSC and TM differences. First, qRT-PCR was performed on RNA isolated from cell lysates. Second, induction of myocilin expression after 500 nM Dex treatment for 1 week was assessed using western blots. Finally, cell contractility was determined by measuring area vs. time in free-floating cell laden collagen I gels.

 
Results
 

Differentially expressed mRNAs between MSCs and TM cells were observed (subset shown in Figure 1 for 4 different TM strains where 2 strains with more juxtacanalicular [JCT] cells are noted), with MGP, MYOC and Plas-Act message levels higher in most TM strains. In addition, JCT/TM strains differentially expressed lower CHI3l1 levels compared to TM-only strains. Dex led to c. 20-fold higher MYOC protein expression in TM cells vs. MSCs. TM cells were found to be less contractile than MSCs over 7 - 10 day time scales.

 
Conclusions
 

qRT-PCR identified a number of markers upregulated in TM cells vs. MSCs. TM cell and MSC strains were found to be heterogeneous in their expression profile, and subsequent studies should use multiple cell strains to account for cell variability. Dex induction of MYOC was confirmed as a robust but not rapid characterization approach. Differences were also noted in contractile properties of the cell types, with MSCs more contractile than TM cells. Next steps will expose MSCs to relevant biophysical stimuli to establish a differentiation protocol to a TM lineage.  

 
Figure 1: mRNA expression (log scale) in TM cells relative to average expression in three MSC lines. (Mean ± SEM; * p < 0.01, ** p < 0.05, *** p < 0.001).
 
Figure 1: mRNA expression (log scale) in TM cells relative to average expression in three MSC lines. (Mean ± SEM; * p < 0.01, ** p < 0.05, *** p < 0.001).

 
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