Neural Progenitor Cells Derived From Human Amniotic Membrane as a Potential Donor Cells for Retinal Regeneration

Abstract

Abstract: : Purpose: Neural stem/progenitor cells are regarded as promising tools for regenerative medicine in the retina. As the source of the cells, somatic stem cells derived from the retina or brain and embryonic stem cells are currently used in experimental animals. In humans, however, such cells are accompanied with significant problems concerning ethics, supply in quantity, and immunological rejection. If human amniotic membrane can be the source of the stem cells, the involvement with these issues should be avoided. We investigated a potential of human amniotic epithelial (HAE) cells as a donor for neural progenitor cells. Methods: Human amniotic membrane was mechanically peeled from chorion of a placenta obtained from an uncomplicated elective Caesarian section with the informed consent of each donor. The amniotic membrane was treated with trypsin, and then HAE cells were harvested. The HAE cells were cultured with various media, and the expression of markers for neural progenitors and mature neural cells was assessed with RT-PCR and immunocytostaining. Results: The cultured HAE cells showed typical neural progenitor cell-like morphology with round shape, and expressed nestin and Musashi, makers for neural progenitors. After 3weeks of the cultivation with neurobasal medium supplemented with epidermal growth factor, these cells extended neurite-like processes and showed network-formation. In these cells, nestin expression was reduced and the expression of microtubule-associated protein 2, a marker for neurons, was upregulated. Conclusions: Neural progenitor cells were obtained from human amniotic membrane. It is suggested that human amniotic membrane may be usable as a source of neural progenitor cells for regenerative medicine in the central nervous system, including the retina.

Keywords: regeneration • retina • transplantation