Similarly derived and cultured hESC lines show variation in their developmental potential towards neuronal cells in long-term culture.
BACKGROUND
Human embryonic stem cells (hESCs) can differentiate into any human cell type, including CNS cells, and thus have high potential in regenerative medicine. Several protocols exist for neuronal differentiation of hESCs, which do not necessarily work for all hESC lines.


METHODS
We tested the differentiation capacity of four similarly derived and cultured hESC lines (HS181, HS360, HS362 and HS401) in suspension culture in relatively simple neural differentiation medium for up to 20 weeks.


RESULTS
All the hESC lines differentiated into neuronal cells, but in a line-dependent manner. Using our method, the HS181- and HS360-derived neurospheres differentiated in vitro into pure neuronal cell populations within 6 weeks, whereas HS362 and HS401 reached their peak of differentiation in 12 weeks, but never produced pure neuronal cell populations using the present method. The withdrawal of FGF from suspension culture increased the in vitro differentiation potential. The hESC-derived neurospheres formed functional neuronal networks when replated on a microelectrode array and responded as expected to pharmacologic modulation.


CONCLUSIONS
Simple neurosphere culture is a suitable method for producing hESC-derived neuronal cells that can form functional neuronal networks from a number of hESC lines. The variation in the differentiation potential of hESC lines into neuronal cells must be carefully considered by those comparing various differentiation methods and designing transplantation therapies for neuronal disorders.
Lappalainen
Riikka S
RS
Regea Institute for Regenerative Medicine, University of Tampere & Tampere University Hospital, Tampere, Finland.
Salomäki
Minna
M
Ylä-Outinen
Laura
L
Heikkilä
Teemu J
TJ
Hyttinen
Jari A K
JA
Pihlajamäki
Harri
H
Suuronen
Riitta
R
Skottman
Heli
H
Narkilahti
Susanna
S
D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019070:Cell Lineage; D003470:Culture Media / Q000737:chemistry; D053595:Embryonic Stem Cells / Q000166:cytology*; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D006801:Humans; D009474:Neurons / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000166:cytology; D044968:Regenerative Medicine / Q000379:methods; D013997:Time Factors
D003470:Culture Media; D005346:Fibroblast Growth Factors