March 2012
Volume 53, Issue 14
Free
ARVO Annual Meeting Abstract  |   March 2012
Co-culturing Adipose-derived Stem Cells (ASCs) with Adult Keratocytes, a Novel Method of Differentiation
Author Affiliations & Notes
  • Ladan Espandar
    Ophthalmology,
    Tulane University, New Orleans, Louisiana
  • Shijia Zhang
    Center for Stem Cell Research and Regenerative Medicine,
    Tulane University, New Orleans, Louisiana
  • Kathleen M. Imhof
    Department of Cell & Molecular Biology,
    Tulane University, New Orleans, Louisiana
  • Bruce A. Bunnell
    Center for Stem Cell Research and Regenerative Medicine,
    Tulane University, New Orleans, Louisiana
  • Footnotes
    Commercial Relationships  Ladan Espandar, None; Shijia Zhang, None; Kathleen M. Imhof, None; Bruce A. Bunnell, None
  • Footnotes
    Support  CTRECP GRANT
Investigative Ophthalmology & Visual Science March 2012, Vol.53, 3506. doi:https://doi.org/
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    • Get Citation

      Ladan Espandar, Shijia Zhang, Kathleen M. Imhof, Bruce A. Bunnell; Co-culturing Adipose-derived Stem Cells (ASCs) with Adult Keratocytes, a Novel Method of Differentiation. Invest. Ophthalmol. Vis. Sci. 2012;53(14):3506. doi: https://doi.org/.

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

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Abstract
 
Purpose:
 

To evaluate the efficiency of co-culture system for ASCs to differentiate to keratocytes.

 
Methods:
 

Co-culture Methods: Transwell inserts with 0.4-micron pores were placed into the fibronectin-coated 6-well plates that contained passage 3~4 of h-ASCs in basal keratocyte differentiation medium (KDM); and human keratocytes were inoculated into the inserts in KDM medium. The cells were allowed to culture for 14 days before RNA isolation.Control Groups: Cultured keratocytes in KDM medium and cultured h-ASCs in KDM and complete culture medium (CCM) were used as control groups for co-cultured keratocyts and h-ASCs respectively.q-RT PCR: After two weeks of cell culturing, total RNA was isolated using an RNeasy mini kit. The real-time PCR reactions were performed as previously described with slight modification. Human keratocan and aldehyde dehydrogenase (ALDH3A1) mRNA levels were assayed using Taq-Man Gene Expression Assays.

 
Results:
 

q-RT PCR was performed for ALDH3A1 and human keratocan in h-ASCs and keratocytes in co-cultured and control groups separately. Results are shown in Figure 1 A, B and Figure 2 A, B.

 
Conclusions:
 

Co-culture system may enhance and facilitate differentiation of h-ASCs. There is an absolute need for more studies to explore interaction between stem cells and adult cells in co-culture system.  

 

 
Keywords: cornea: stroma and keratocytes • cornea: basic science 
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