Abstract
Purpose: :
The induced pluripotent stem (iPS) cell is expected as a powerful tool for research of regenerative medicine. Previously we reported that monkey embryonic stem (ES) cell-derived retinal pigment epithelial (RPE) cells rescued photoreceptor cells after transplantation into RCS rats (Haruta et al. IOVS, 2004). In this report we produced iPS cell lines from monkeys to investigate their ability to differentiate into retinal cells.
Methods: :
The fibroblasts derived from cynomolgus monkey abdominal skin were infected with retroviruses carrying oct3/4, sox2, klf4 and c-myc genes. They were cultured in ES cell maintaining medium on STO feeder cells. Next, the established monkey iPS cells were induced to differentiate into RPE cells by culturing with the supernatant of PA6, a mouse stromal cell line. The property of the differentiated RPE cells were analyzed.
Results: :
After one month from retroviral infection, some colonies appeared among the fibroblasts. The colonies were picked up and expanded as cell lines. These cell lines showed monkey ES cell-like morphology and expressed ES cell markers such as alkaline phosphatase, Nanog and SSEA-4. By making teratoma in SCID mice, these cells were confirmed to have the ability to differentiate into three germ layers. In addition, these iPS cells had capability of differentiating into retinal cells including RPE cells with pigment and polygonal shapes. These cells expressed PRE cell-specific markers and exhibited phagocytotic function in vitro.
Conclusions: :
We established iPS cells lines from monkey skin fibroblast. The RPE cells derived from these monkey iPS cells can be used for autologous or allogenic transplantation to evaluate the immune rejection and their function in vivo.
Keywords: regeneration • retinal pigment epithelium • differentiation