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Journal of Materials Science: Materials in Medicine

, Volume 24, Issue 12, pp 2819–2829 | Cite as

Chitosan–gelatin biopolymers as carrier substrata for limbal epithelial stem cells

  • Ana de la Mata
  • Teresa Nieto-Miguel
  • Marina López-Paniagua
  • Sara Galindo
  • María Rosa Aguilar
  • Luis García-Fernández
  • Sandra Gonzalo
  • Blanca Vázquez
  • Julio San Román
  • Rosa María Corrales
  • Margarita Calonge
Article

Abstract

The aim of this work was to evaluate semi-synthetic biopolymers based on chitosan (CH) and gelatin (G) as potential in vitro carrier substrata for human limbal epithelial cells (hLECs). To that end, human corneal epithelial cells (HCE) were cultured onto different CH–G membranes. None of the polymers were cytotoxic and cell proliferation was higher when CH was functionalized with G. Expression levels of corneal epithelial markers (K3, K12, E-caherin, desmoplakin, and zonula occludens (ZO)-1) were better maintained in HCE cells grown on CH–G 20:80 membranes than other proportions. Consequently, CH–G 20:80 was chosen for the subsequent expansion of hLECs. Cells derived from limbal explants were successfully expanded on CH–G 20:80 membranes using a culture medium lacking components of non-human animal origin. The expression levels found for corneal (K3 and K12) and limbal epithelial stem cells (K15) specific markers were similar to or higher than those found in limbal cells grown onto the control substratum. Our results demonstrate that CH–G 20:80 membranes are suitable for the expansion and maintenance of stem cells derived from the limbal niche. These results strongly support the use of polymers as alternative substrata for the transplantation of cultivated limbal cells onto the ocular surface.

Keywords

Chitosan Ocular Surface Tissue Culture Plastic Corneal Epithelial Cell Real Time Reverse Transcriptase Polymerase Chain Reaction 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This work has been supported by the Instituto de Salud Carlos III (CIBER-BBN CB06/01/003), Spain. Centro en Red de Medicina Regenerativa y Terapia Celular, Castilla y León, Spain. Junta de Castilla y León, Spain: SAN673/VA/28/08, SAN126/VA11/09. CIBER-BBN is an initiative funded by the VI National R&D&i Plan 2008–2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. S. Galindo and M. López-Paniagua were supported by scholarships co-financed by the Castilla y León Government and the European Social Fond. We thank M. F. de la Paz, MD, (Barraquer Eye Bank of Barcelona, Spain) and J. Iglesias, PhD, (Tissue Establishment, San Francisco Clinic Fundation, Leon, Spain) for providing human corneoscleral tissues. We also thank J. M. Herreras, MD, for his clinical advice, V. Sáez (IOBA laboratory technician) for her initial technical assistance and I. Fernández (IOBA, University of Valladolid, Spain) for statistical assistance We thank S. Gonzalo for her technical support in the polymer characterization experiments. And we thank B. Bromberg (Certified Editor in Life Science of Xenofile Editing, www.xenofileediting.com) for his assistance in the final editing and preparation of this manuscript. Presented in part as an abstract at the 80th Annual meeting of the Association for Research in Vision and Ophtalmology (ARVO), May 2009, Fort Lauderdale, FL, USA (IOVS 2009, ARVO-E-Abstract 6288); at the 22nd European Conference on Biomaterials (ESB2009), September 2009, Lausanne (Switzerland), poster 715 0102; and at the Biofuture 2011: Young European Biomaterial Scientists Designing a View for the Future, November 2011, Gante (Belgium). No competing financial interests exist.

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Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Ana de la Mata
    • 1
    • 2
  • Teresa Nieto-Miguel
    • 2
    • 1
  • Marina López-Paniagua
    • 1
    • 2
  • Sara Galindo
    • 1
    • 2
  • María Rosa Aguilar
    • 2
    • 3
  • Luis García-Fernández
    • 3
    • 4
  • Sandra Gonzalo
    • 2
    • 3
    • 5
  • Blanca Vázquez
    • 2
    • 3
  • Julio San Román
    • 2
    • 3
  • Rosa María Corrales
    • 1
    • 2
    • 6
  • Margarita Calonge
    • 1
    • 2
  1. 1.IOBA (Institute of Applied Ophthalmobiology)University of ValladolidValladolidSpain
  2. 2.CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN)BarcelonaSpain
  3. 3.Institute of Polymer Science and Technology, Spanish National Research Council (CSIC)MadridSpain
  4. 4.Active Surfaces and Materials GroupMax-Planck-Institut für PolymerforschungMainzGermany
  5. 5.Molecular Biology Center Severo Ochoa (CBM)MadridSpain
  6. 6.Ocular Surface CentreBaylor College of MedicineHoustonUSA

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