Self-contained induction of neurons from human embryonic stem cells.
BACKGROUND
Neurons and glial cells can be efficiently induced from mouse embryonic stem (ES) cells in a conditioned medium collected from rat primary-cultured astrocytes (P-ACM). However, the use of rodent primary cells for clinical applications may be hampered by limited supply and risk of contamination with xeno-proteins.


RESULTS
We have developed an alternative method for unimpeded production of human neurons under xeno-free conditions. Initially, neural stem cells in sphere-like clusters were induced from human ES (hES) cells after being cultured in P-ACM under free-floating conditions. The resultant neural stem cells could circumferentially proliferate under subsequent adhesive culture, and selectively differentiate into neurons or astrocytes by changing the medium to P-ACM or G5, respectively. These hES cell-derived neurons and astrocytes could procure functions similar to those of primary cells. Interestingly, a conditioned medium obtained from the hES cell-derived astrocytes (ES-ACM) could successfully be used to substitute P-ACM for induction of neurons. Neurons made by this method could survive in mice brain after xeno-transplantation.


CONCLUSIONS
By inducing astrocytes from hES cells in a chemically defined medium, we could produce human neurons without the use of P-ACM. This self-serving method provides an unlimited source of human neural cells and may facilitate clinical applications of hES cells for neurological diseases.
Okuno
Tsuyoshi
T
Advanced Medical Research Laboratory, Mitsubishi Tanabe Pharma Corporation, Osaka, Japan.
Nakayama
Takashi
T
Konishi
Nae
N
Michibata
Hideo
H
Wakimoto
Koji
K
Suzuki
Yutaka
Y
Nito
Shinji
S
Inaba
Toshio
T
Nakano
Imaharu
I
Muramatsu
Shin-Ichi
S
Takano
Makoto
M
Kondo
Yasushi
Y
Inoue
Nobuo
N
D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D017077:Culture Media, Conditioned; D017931:DNA Primers; D018274:Electroporation; D053595:Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D051379:Mice; D009474:Neurons / Q000166:cytology*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D033581:Stem Cell Transplantation
D017077:Culture Media, Conditioned; D017931:DNA Primers