April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Acellular human corneal matrix sheets seeded with human adipose-derived mesenchymal stem cells integrate functionally in an experimental animal model
Author Affiliations & Notes
  • Jorge Luis Alio del Barrio
    St Thomas' Hospital, London, United Kingdom
    Ophthalmology Department, Ramon y Cajal Hospital, Madrid, Spain
  • Maria P De Miguel
    Cell Engineering Laboratory, IdiPAZ, La Paz Hospital Research Institute, Madrid, Spain
  • Jorge L Alio
    Vissum Ophthalmological Institute, Alicante, Spain
  • Massimo Chiesa
    Cell Engineering Laboratory, IdiPAZ, La Paz Hospital Research Institute, Madrid, Spain
  • Nerea Garagorri
    Tecnalia, San Sebastián, Spain
  • Nerea Garcia
    Tecnalia, San Sebastián, Spain
  • Francisco Arnalich-Montiel
    Ophthalmology Department, Ramon y Cajal Hospital, Madrid, Spain
  • Alejandra Rodríguez
    Vissum Ophthalmological Institute, Alicante, Spain
  • Laurent Bataille
    Vissum Ophthalmological Institute, Alicante, Spain
  • Footnotes
    Commercial Relationships Jorge Alio del Barrio, None; Maria De Miguel, None; Jorge Alio, None; Massimo Chiesa, None; Nerea Garagorri, None; Nerea Garcia, None; Francisco Arnalich-Montiel, None; Alejandra Rodríguez, None; Laurent Bataille, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 5158. doi:
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    • Get Citation

      Jorge Luis Alio del Barrio, Maria P De Miguel, Jorge L Alio, Massimo Chiesa, Nerea Garagorri, Nerea Garcia, Francisco Arnalich-Montiel, Alejandra Rodríguez, Laurent Bataille; Acellular human corneal matrix sheets seeded with human adipose-derived mesenchymal stem cells integrate functionally in an experimental animal model. Invest. Ophthalmol. Vis. Sci. 2014;55(13):5158.

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

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Abstract
 
Purpose
 

To evaluate the in vivo biocompatibility of grafts composed of sheets of decellularized human corneal stroma with or without subsequent human adipose derived adult stem cells (h-ADASC) recellularization into the rabbit cornea.

 
Methods
 

Sheets of human corneal stroma of 90 µm thickness were decellularized. Their lack of cytotoxicity was assayed. Recellularization was achieved by the injection, in vitro, of 200 x 103 previously labeled h-ADASC inside the graft and five days of cell culture. The grafts were implanted at 50% depth stromal pocket in vivo and after a controlled and triple masking three-month follow up the animals were sacrificed and the biointegration of the graft, the viability of the stem cells and the expression of keratocan (human keratocyte-specific protein) were assessed.

 
Results
 

Decellularized stromal sheets showed an intact extracellular matrix with a decellularization rate of 92,8% and an excellent recellularization capacity in vitro with h-ADASC. A complete and stable transparency of the graft was observed during the full follow-up with an absence of any clinical sign of inflammation or rejection, which was confirmed by the histology showing no sign of inflammatory infiltrate or just isolated cells. The postmortem analysis demonstrated the survival of the transplanted human stem cells inside the graft and their differentiation into functional human keratocytes detected by the expression of human keratocan.

 
Conclusions
 

We report a new model of lamellar keratoplasty that requires only a simple and safe procedure of liposuction and a donor allogeneic untransplantable cornea to provide an optically transparent autologous stromal graft with demonstrated excellent biocompatibility and integration into the host tissue in a rabbit model.

 
 
The graft remains totally transparent after 12 weeks of follow-up (arrows point to the slightly visible edge of the graft)
 
The graft remains totally transparent after 12 weeks of follow-up (arrows point to the slightly visible edge of the graft)
 
 
A:Hematoxylin-eosin staining of rabbit cornea with an implanted graft of decellularizated human corneal stroma with h-ADASC colonization: hypocellular band of extracellular matrix without vessels or any inflammatory sign (magnification 200X); B: Vybrant CM-DiI positive human cells around and inside the implant (arrows), which express (C) human keratocan (magnification 400X) (Epi: epithelium; Str: stroma)
 
A:Hematoxylin-eosin staining of rabbit cornea with an implanted graft of decellularizated human corneal stroma with h-ADASC colonization: hypocellular band of extracellular matrix without vessels or any inflammatory sign (magnification 200X); B: Vybrant CM-DiI positive human cells around and inside the implant (arrows), which express (C) human keratocan (magnification 400X) (Epi: epithelium; Str: stroma)
 
Keywords: 480 cornea: basic science • 484 cornea: stroma and keratocytes • 741 transplantation  
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