May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Similarities Observed Between Human Limbal and Immature Dental Pulp Stem Cells
Author Affiliations & Notes
  • B. G. Monteiro
    Butantan Institute - USP, Sao Paulo, Brazil
    Genetic Biotechnology, Morphology,
  • R. C. Serafim
    Genetic Biotechnology, Morphology,
    Unifesp, Sao Paulo, Brazil
  • C. M. C. Maranduba
    Butantan Institute - USP, Sao Paulo, Brazil
    Genetic Biotechnology, Morphology,
  • N. F. Lizier
    Genetic Biotechnology, Morphology,
    Unifesp, Sao Paulo, Brazil
  • M. C. P. Silva
    Genetic Biotechnology, Morphology,
    Unifesp, Sao Paulo, Brazil
  • R. L. Smith
    Genetic Biotechnology, Morphology,
    Unifesp, Sao Paulo, Brazil
  • H. Cerruti, II
    CERA Clinic, Sao Paulo, Brazil
  • A. Kerkis
    Clinic and Center of Research in Human Reproduction, Sao Paulo, Brazil
  • J. A. P. Gomes
    Genetic, Ophtalmology,
    Unifesp, Sao Paulo, Brazil
  • I. Kerkis
    Butantan Institute - USP, Sao Paulo, Brazil
    Genetic, Ophtalmology,
  • Footnotes
    Commercial Relationships  B.G. Monteiro, None; R.C. Serafim, None; C.M.C. Maranduba, None; N.F. Lizier, None; M.C.P. Silva, None; R.L. Smith, None; H. Cerruti, CERA Clinic, P; A. Kerkis, None; J.A.P. Gomes, None; I. Kerkis, Butantan Institute, P.
  • Footnotes
    Support  CERA Clinic, FAPESP, CAPES and CNPq
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 6072. doi:https://doi.org/
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    • Get Citation

      B. G. Monteiro, R. C. Serafim, C. M. C. Maranduba, N. F. Lizier, M. C. P. Silva, R. L. Smith, H. Cerruti, II, A. Kerkis, J. A. P. Gomes, I. Kerkis; Similarities Observed Between Human Limbal and Immature Dental Pulp Stem Cells. Invest. Ophthalmol. Vis. Sci. 2008;49(13):6072. doi: https://doi.org/.

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

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Abstract

Purpose: : Expression profile study of human Limbal Stem Cells (hLSC) markers in undifferentiated Human Immature Dental Pulp Stem Cells (hIDPSC) and their implication with corneal epithelium reconstruction in animal model with Limbal Stem Cells Deficiency (LSCD).

Methods: : hIDPSC were cultivated according to Kerkis et al. (2006). The expression profile of hIDPSC was studied using markers for limbal stem cells (ABCG2), integrin beta-1, conexin 43, p63 and corneal epithelium (keratin-3). The expression was analyzed by immunocitochemistry and RT-PCR. Moreover, the expression of epithelial and corneal stem cells markers were evaluated in vivo after hIDPSC transplantation in LSCD model, which was induced by chemically burn in rabbits. Control animals received only amniotic membrane. Corneal epithelium formed onto the rabbit eyes were submitted to immunohistochemistry and RT-PCR analyses, with the same aforementioned markers besides histological study. Transmission electron microscopy (TEM) was also performed.

Results: : RT-PCR results showed that undifferentiated hIDPSC and LSC present similar pattern of epithelial expression as well as LSC markers. After hIDPSC transplantation into injured rabbit eye, the transparency of the cornea was improved indicating differentiation of hIDPSC into corneal tissue, which was demonstrated by immunohistochemistry. The use of human specific antibodies against p63 and integrin β1 confirmed the hIDPSC contribution into new formed tissue, which reacted positively with K3, a specific antibody against cornea. In control group, the conjunctiva fully covered the cornea and was strongly vascularized, while reacting negatively for the markers. Histological and TEM analyses confirmed the formation of corneal epithelium, while in control group the cornea structure was chaotically organized.

Conclusions: : Our data showed hIDPSC present gene expression profile similar to those observed in hLSC. Moreover, after their transplantation into the injured rabbit eye they differentiated and presented human cornea gene expression pattern as well as formed corneal epithelium. These similarities between hIDPSC and hLSC suggest these cells are a strong candidate to be used in corneal epithelium reconstruction, which can solve the problem of donor hLSC in TLD case.

Clinical Trial: : http://www.butantan.gov.br/ceuaib/index.htm

Keywords: cornea: basic science • gene/expression • cornea: clinical science 
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