An improved protocol that induces human embryonic stem cells to differentiate into neural cells in vitro.
Human embryonic stem (ES) cells have the capacity for self-renewal and are able to differentiate into any cell type. However, obtaining high-efficient neural differentiation from human ES cells remains a challenge. This study describes an improved 4-stage protocol to induce a human ES cell line derived from a Chinese population to differentiate into neural cells. At the first stage, embryonic bodies (EBs) were formed in a chemically-defined neural inducing medium rather than in traditional serum or serum-replacement medium. At the second stage, rosette-like structures were formed. At the third stage, the rosette-like structures were manually selected rather than enzymatically digested to form floating neurospheres. At the fourth stage, the neurospheres were further differentiated into neurons. The results show that, at the second stage, the rate of the formation of rosette-like structures from EBs induced by noggin was 88+/-6.32%, higher than that of retinoic acid 55+/-5.27%. Immunocytochemistry staining was used to confirm the neural identity of the cells. These results show a major improvement in obtaining efficient neural differentiation of human ES cells.
Zhou
Jun-Mei
JM
Center for Developmental Biology, Xinhua Hospital, School of Medicine, Shanghai Jiao Tong University, 1665 Kong Jian Road, Shanghai 200092, PR China.
Chu
Jian-Xin
JX
Chen
Xue-Jin
XJ
D044466:Asian People; D002352:Carrier Proteins / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002681:China; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D009474:Neurons / Q000166:cytology*; D014212:Tretinoin / Q000494:pharmacology; D000099176:Noggin Protein
D002352:Carrier Proteins; D014212:Tretinoin; D000099176:Noggin Protein