June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Expression of Pluripotent Markers L1CAM and SSEA-5, Common to Human Retinoblastomas, Xenografts, Teratomas, Embryonic Tumors and Induced Pluripotent Stem Cells
Author Affiliations & Notes
  • Gail Seigel
    Center for Hearing and Deafness, University at Buffalo, Buffalo, NY
    SUNY Eye Institute, Buffalo, NY
  • Meerim Choi
    Center for Hearing and Deafness, University at Buffalo, Buffalo, NY
    SUNY Eye Institute, Buffalo, NY
  • Rui Chang
    Genetics and Genomic Sciences, Mount Sinai School of Medicine, New York, NY
  • Jason Meyer
    Biology, Indiana University, Indianapolis, IN
  • Bruce Ksander
    Ophthalmology, Schepens Eye Research Institute, Boston, MA
  • Paraskevi Kolovou
    Ophthalmology, Schepens Eye Research Institute, Boston, MA
  • Nadine de Waard
    Ophthalmology, Schepens Eye Research Institute, Boston, MA
    Ophthalmology, Leiden University, Leiden, Netherlands
  • Linda Cassidy
    Center for Hearing and Deafness, University at Buffalo, Buffalo, NY
    SUNY Eye Institute, Buffalo, NY
  • Footnotes
    Commercial Relationships Gail Seigel, None; Meerim Choi, None; Rui Chang, None; Jason Meyer, None; Bruce Ksander, None; Paraskevi Kolovou, None; Nadine de Waard, None; Linda Cassidy, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 1255. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Gail Seigel, Meerim Choi, Rui Chang, Jason Meyer, Bruce Ksander, Paraskevi Kolovou, Nadine de Waard, Linda Cassidy; Expression of Pluripotent Markers L1CAM and SSEA-5, Common to Human Retinoblastomas, Xenografts, Teratomas, Embryonic Tumors and Induced Pluripotent Stem Cells. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1255.

      Download citation file:


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

      ×
  • Supplements
Abstract
 
Purpose
 

Cancer stem cells and therapeutic stem cells share common and divergent pathways involved in cell growth and differentiation. In this study, we sought to bridge the gap between retinoblastoma (RB) tumor cells and iPSCs undergoing retinal differentiation by testing the hypothesis that L1CAM and SSEA-5 could be used to evaluate pluripotency in both models.

 
Methods
 

Magnetically enriched stem-like ABCG2+ and non-stem like ABCG2- cells from the human RB143 retinoblastoma cell line underwent microarray analysis using the Agilent platform. We used ensemble based statistical models to predict differentially expressed genes of pluripotency. The larger list was condensed into genes with biological relevance to the behavior of cancer stem cells and iPSCs. One of these genes was L1CAM, a neural cell adhesion marker. We included SSEA-5 for analysis based on promising published data as a novel marker of pluripotency. We used immunohistochemistry to analyze RB tumors and cell lines, RB xenografts, human embryonic tumors, iPSC-derived teratomas, as well as iPS cells before and after previously established retinal differentiation protocols to determine whether these markers would be useful for monitoring pluripotency. The expression of L1CAM in these cells was also assessed by qRT-PCR.

 
Results
 

RB143 cells and xenografts were immunoreactive to L1CAM and SSEA-5. L1CAM was expressed in most human RB and human embryonic tumors examined. SSEA-5 was preferentially expressed in embryonic tumors and some RB tumors. In iPS-induced teratomas, we saw immunoreactivity to both markers, with some areas of co-localization. Undifferentiated iPSCs exhibited strong immunoreactivity to SSEA-5 and L1CAM, with a few positive cells remaining even after 20 days of retinal differentiating treatment. L1CAM showed little change by qRT-PCR, suggesting post-transcriptional regulation.

 
Conclusions
 

Both SSEA-5 and L1CAM are cell surface markers that show promise as indicators of pluripotency. Stem cell regulatory mechanisms common to RB cancer stem cells and retinal therapeutic stem cells may be useful in monitoring and controlling the growth and differentiation of iPSCs. Further studies may aid in growth regulatory strategies that are essential to the development of safe ocular stem cell replacement therapies.

 
 
Embryonal carcinoma: SSEA-5 (red), L1CAM (green)
 
Embryonal carcinoma: SSEA-5 (red), L1CAM (green)
 
Keywords: 703 retinoblastoma • 721 stem cells • 744 tumors  
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×