CoCl(2) inhibits neural differentiation of retinoic acid-treated embryoid bodies.
The effects of CoCl(2) on retinoic acid (RA)-treated embryoid bodies (EBs) were investigated. Four-day EBs were treated with 5x10(-6) M of RA for 4 d, then subjected to attached culturing for 7 d in the presence of CoCl(2) at 0, 20, and 100 microM. Differentiation into MAP2- and GFAP-immunopositive cells was inhibited by CoCl(2) in a dose-dependent manner. Next, RA-treated EBs were dissociated into single cells and cultured for 7 d at an initial cell density of 1x10(3)/cm(2). The number of cells increased in a CoCl(2)-dose dependent fashion. In cultures with 100 microM of CoCl(2), more than 90% of the cells were immunopositive for nestin and nestin-immunopositive cells formed clusters, while there were few cells immunopositive for MAP2 or GFAP. These results suggest that CoCl(2) inhibits neural differentiation of RA-treated EB cells and promotes the proliferation of nestin-immunopositive cells, i.e., embryonic stem (ES)-derived neural stem-like cells.
Nonaka
Jun-ichi
J
Department of Neurosurgery, Nara Medical University, 840 Shijo-cho, Kashihara, Nara, Japan.
Yoshikawa
Masahide
M
Ouji
Yukiteru
Y
Matsuda
Ryosuke
R
Nishimura
Fumihiko
F
Yamada
Shuichi
S
Nakase
Hiroyuki
H
Moriya
Kei
K
Nishiofuku
Mariko
M
Ishizaka
Shigeaki
S
Sakaki
Toshisuke
T
D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation; D003035:Cobalt / Q000494:pharmacology*; D017931:DNA Primers; D004305:Dose-Response Relationship, Drug; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects*; D007150:Immunohistochemistry; D009474:Neurons / Q000166:cytology / Q000187:drug effects*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014212:Tretinoin / Q000494:pharmacology*
D017931:DNA Primers; D003035:Cobalt; D014212:Tretinoin; C018021:cobaltous chloride