Cyclopamine treatment of human embryonic stem cells followed by culture in human astrocyte medium promotes differentiation into nestin- and GFAP-expressing astrocytic lineage.
Human embryonic stem cells (hESCs) are able to differentiate into various cell types, including neuronal cells and glial cells. However, little information is available regarding astrocyte differentiation. This report describes the differentiation of hESCs into nestin- and GFAP-expressing astrocytes following treatment with cyclopamine, which is an inhibitor of Hedgehog (Hh) signaling, and culturing in human astrocyte medium (HAM). In hESCs, cyclopamine treatment suppressed the expression of Hh signaling molecules, the Hh signaling target gene, and ESC-specific markers. Clyclopamine also induced the differentiation of the cells at the edges of the hESC colonies, and these cells stained positively for the early neural marker nestin. Subsequent culturing in HAM promoted the expression of the astrocyte-specific marker GFAP, and these cells were also nestin-positive. These findings indicate that treatment with cyclopamine followed by culturing in HAM leads to the differentiation of hESCs into nestin- and GFAP-expressing astrocytic lineage.
Lee
Dong-Seok
DS
Laboratory of Human Genomics, Korea Research Institute of Bioscience and Biotechnology, Daejeon 300-303, Republic of Korea.
Yu
Kweon
K
Rho
Jeung-Yon
JY
Lee
Eunyoung
E
Han
Jee-Soo
JS
Koo
Deog-Bon
DB
Cho
Yee Sook
YS
Kim
Janghwan
J
Lee
Kyung-Kwang
KK
Han
Yong-Mahn
YM
D001253:Astrocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D019070:Cell Lineage / Q000187:drug effects*; D017077:Culture Media, Conditioned; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000096:biosynthesis*; D053823:Hedgehog Proteins / Q000037:antagonists & inhibitors / Q000235:genetics; D006801:Humans; D007381:Intermediate Filament Proteins / Q000096:biosynthesis*; D009419:Nerve Tissue Proteins / Q000096:biosynthesis*; D064231:Nestin; D009457:Neuroglia / Q000187:drug effects / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D014704:Veratrum Alkaloids / Q000494:pharmacology*
D017077:Culture Media, Conditioned; D005904:Glial Fibrillary Acidic Protein; D053823:Hedgehog Proteins; D007381:Intermediate Filament Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; D014704:Veratrum Alkaloids; C000541:cyclopamine