Duration 5min, Q&A 3min
Transplantation of 3-Dimensional ESC and iPSC-derived Retinal-Like Sheet to Retinal Degenerative Mice
To evaluate for the competency of these 3D differentiated ESC- or iPSC-derived retinal sheets for their potency as a graft for therapeutic transplantation.
Mouse ES cell line (Rx-GFP knocked-in)
Mouse iPS cell line (Nrl-GFP transgenic)
6-8 wks C3H/HejYokSlc (rd1 mice)
The mouse ES cell line and mouse iPS cell line were differentiated to stratified neural retina by a modified method of Eiraku's (Nature, 2011). The developmental stage of the differentiated cultures was determined by immunostaining in comparison with the developing mouse retina. Next, the mESC- or miPSC-derived retinal tissues were transplanted into the subretinal space of rd1 retina. Their survival, maturation and integration were immunohistologically evaluated.
Results and Conclusion
The neural retina progenitor induction were increased by modifying the method by Eiraku. Both mESC- and miPSC-derived retinal tissue expressed Recoverin, Crx, Calretinin and Rhodopsin in the similar pattern to those of the developing mouse retina. The DD.21 was approximately equivalent to postnatal day1 (P1) and cultured tissues less than DD.20 were considered comparable to embryonic retinal tissue. Both mESC and iPSC derived retinal tissues transplanted to subretinal space of rd1, could survive and developed mature photoreceptors with the formation of variable degree of IS/OS in degenerating environment. About 83% (35/42) of the differentiated tissue of younger than DD.18 at the time of transplantation retained structured ONL after transplantation. Some of these grafts with structured ONL showed direct contact with the host retina with possible synaptic connection showed by synaptic marker immunostaining. We further identified ribbon synapses in the area of host-graft interphase. The three-dimensional retina-like sheets derived from Rx-GFP mESC and Nrl-GFP miPSC could serve as grafts in transplantation therapy for retinal degeneration.
[ Keyword ]
Retina / Embryonic stem cells / Induced pluripotent stem cells / Retinal transplantation / Retinal differentiation
[ Conflict of Interest ]