Abstract
Regenerative medicine in ophthalmology that uses induced pluripotent stem cells (iPS) cells has been described, but those studies used iPS cells derived from fibroblasts. Here, we generated iPS cells derived from iris cells that develop from the same inner layer of the optic cup as the retina, to regenerate retinal nerves. We first identified cells positive for p75NTR, a marker of retinal tissue stem and progenitor cells, in human iris tissue. We then reprogrammed the cultured p75NTR-positive iris tissue stem/progenitor (H-iris stem/progenitor) cells to create iris-derived iPS (H-iris iPS) cells for the first time. These cells were positive for iPS cell markers and showed pluripotency to differentiate into three germ layers. When H-iris iPS cells were pre-differentiated into neural stem/progenitor cells, not all cells became positive for neural stem/progenitor and nerve cell markers. When these cells were pre-differentiated into neural stem/progenitor cells, sorted with p75NTR, and used as a medium for differentiating into retinal nerve cells, the cells differentiated into Recoverin-positive cells with electrophysiological functions. In a different medium, H-iris iPS cells differentiated into retinal ganglion cell marker-positive cells with electrophysiological functions. This is the first demonstration of H-iris iPS cells differentiating into retinal neurons that function physiologically as neurons.
Highlights
IntroductionThe human body is composed of approximately 3.72 × 1013 cells of about 270 types, as estimated in 2013 by Bianconi et al using bibliographical and/or mathematical approaches [1]
We created human iris-derived induced pluripotent stem cells (iPS) (H-iris iPS) cells for the first time by using iris tissue stem cells (H-iris stem/progenitor cells) selected by p75NTR, and we examined the differentiation of these iPS cells into Recoverin-positive cells expressing Recoverin and ganglion cells expressing Neurofilament-M and Brn-3b
H-iris iPS cells were created from H-iris stem/progenitor cells
Summary
The human body is composed of approximately 3.72 × 1013 cells of about 270 types, as estimated in 2013 by Bianconi et al using bibliographical and/or mathematical approaches [1]. Until 2000, the basic research conducted to differentiate these various cell types in vitro used mainly embryonic stem (ES) cells (Table 1).
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