The biological characteristics of amniotic membrane and it's application for regenerative medicine.

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1) Amniotic epithelial cell, mesenchymal cell and umbilical mesenchymal cell, cultured under ultra low attached dish, made spheres that had self-renewal ability, expression of mRNA or proteins of stem cell markers (Nanog, Oct3/4, SSEA4).2) We made 11 immortalized cell lines of amniotic epithelial cell, mesenchymal cell, umbilical mesenchymal cells, which work were collaborated with Prof. Kiyono (National Cancer Center). Those cell lines were made by virus-transfection of gene such as E6E7, hTERT, bmil and cyclin inhibitor. We selected some cell lines that did not change cell growth and morphology even though over 50 PDL from these 11 lines. Those selected cell lines differentiated into bone, cartilage and adipose cells.3) Dried amniotic membrane were used for operation of the pterygium and aqueous humor tunnel plasty under the glaucoma. Under these treatment, shortening of the operative time, available for the large tissue defect were shown as advantages.4) i) The epithelial surface of dried amniotic membrane suppress for adhesion with the tissue, and stromal surface adherenced to tissue. And the dried amniotic membrane worked as a good scaffold even if the surface was epithelium. The conjunctiva cells grew and multiplied on the dried amniotic membrane which we transplanted in the lesion. ii) It was easy to handle and make the dried amniotic membrane for the suitable size and shape that it was compared with fresh or cryopreserved amniotic membrane, and wrinkles were hard to occur. These advantages become that an ophthalmologist could be short the operative time, could be available for the large tissue defect. iii) We developed a polymer to solubilize hydrophobic antiviral medication for treatment keratitis herpetic. We impregnated the solubilized antiviral medication to dried amniotic membrane, and presented with a similar effect that of treatment with the ointment in rabbit model.