Expansion and neural differentiation of embryonic stem cells in adherent and suspension cultures.
The embryonic stem cell line, S25, is a genetically modified line that allows lineage selection of neural cells (M. Li, L. Lovell-Badge, A. Smith (1998) Current Biology 8: 971-974). Here, the growth parameters of this cell line were analysed. Serial passaging in adherent conditions enabled these cells to grow rapidly (average specific growth rates of 0.035 h-1) and generate high viable cell densities (above 90%). The aggregation of the S25 cells into embryoid bodies (EBs) was also studied, indicating limited cell growth (maximum cell densities of 2.7 x 10(5) cells ml-1) and a high variability of aggregate size (70-400 microns after 8 d). Enzymatic dissociation of EBs with 1% (v/v) trypsin gave highest cell viability (91%) and density (1.4 x 10(4) cells ml-1) and the cells thus obtained are able to differentiate into neurons.
Abranches
Elsa
E
Centro de Engenharia Biológica e Química, Instituto Superior Técnico, Faculdade de Medicina de Lisboa, Lisboa, Portugal. elsa.abranches@popsrv.ist.utl.pt
Bekman
Evguenia
E
Henrique
Domingos
D
Cabral
Joaquim M S
JM
Albranches
Elsa
E
D000818:Animals; D002448:Cell Adhesion / Q000502:physiology; D002449:Cell Aggregation / Q000502:physiology; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D051379:Mice; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D014357:Trypsin / Q000494:pharmacology
D014357:Trypsin