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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)
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).
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.
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.
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.
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