Self-formation of optic cups and storable stratified neural retina from human ESCs.
In this report, we demonstrate that an optic cup structure can form by self-organization in human ESC culture. The human ESC-derived optic cup is much larger than the mouse ESC-derived one, presumably reflecting the species differences. The neural retina in human ESC culture is thick and spontaneously curves in an apically convex manner, which is not seen in mouse ESC culture. In addition, human ESC-derived neural retina grows into multilayered tissue containing both rods and cones, whereas cone differentiation is rare in mouse ESC culture. The accumulation of photoreceptors in human ESC culture can be greatly accelerated by Notch inhibition. In addition, we show that an optimized vitrification method enables en bloc cryopreservation of stratified neural retina of human origin. This storage method at an intermediate step during the time-consuming differentiation process provides a versatile solution for quality control in large-scale preparation of clinical-grade retinal tissues.
Organogenesis and Neurogenesis Group, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan; Division of Human Stem Cell Technology, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan; Environmental Health Science Laboratory, Sumitomo Chemical Co., Ltd., Osaka 554-8558, Japan; Department of Medical Embryology, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan.
Tokushige
T
Nakano
Organogenesis and Neurogenesis Group, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan; Division of Human Stem Cell Technology, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan; Environmental Health Science Laboratory, Sumitomo Chemical Co., Ltd., Osaka 554-8558, Japan.
Satoshi
S
Ando
Organogenesis and Neurogenesis Group, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan.
Nozomu
N
Takata
Organogenesis and Neurogenesis Group, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan.
Masako
M
Kawada
Organogenesis and Neurogenesis Group, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan.
Keiko
K
Muguruma
Laboratory of Extracellular Matrix Biochemistry, Institute for Protein Research, Osaka University, Suita 565-0871, Japan.
Kiyotoshi
K
Sekiguchi
Environmental Health Science Laboratory, Sumitomo Chemical Co., Ltd., Osaka 554-8558, Japan.
Koichi
K
Saito
Electron Microscopy Laboratory, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan.
Shigenobu
S
Yonemura
Organogenesis and Neurogenesis Group, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan; Division of Human Stem Cell Technology, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan.
Mototsugu
M
Eiraku
Organogenesis and Neurogenesis Group, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan; Division of Human Stem Cell Technology, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan; Department of Medical Embryology, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan. Electronic address: yoshikisasai@cdb.riken.jp.
Yoshiki
Y
Sasai
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology*; D005123:Eye / Q000196:embryology*; D006801:Humans; D051379:Mice; D038081:Organogenesis; D044968:Regenerative Medicine; D055351:Retinal Neurons / Q000166:cytology*