Significant variations in differentiation properties between independent mouse ES cell lines cultured under defined conditions.
Mouse embryonic stem (ES) cells are isolated from the inner cell mass (ICM)/epiblast of preimplantation embryos and are widely used in cell differentiation studies. We have previously observed differences in transcript and antigen expression following differentiation of ES cells lines in vitro. We have investigated this further by comparing the differentiation characteristics of five independently derived ES cell lines cultured and differentiated under defined conditions. Undifferentiated ES cell lines exhibited similar morphology and antigen/transcript marker expression. However, upon differentiation in monolayer culture by LIF withdrawal, only two of the lines expressed similar germ layer transcript profiles, and these were significantly altered compared to differentiation in serum-supplemented media. Neurofilament-68k was the only transcript marker common to all cell lines, however, induction of neuroectoderm lineages using 1 microM all-trans retinoic acid (RA) resulted in significant variations in cell number and morphology between the lines. Furthermore, neurons were only formed from clones of the two cell lines that exhibited similar transcript profiles, although the morphology was different between the two. We conclude that the independent ES cell lines in this study differ in their response to alterations in culture conditions in vitro, and the use of an appropriate cell line enables relatively homogeneous neuronal populations to be achieved in monolayer culture under defined conditions.
Ward
Christopher M
CM
Immunology Group, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Wilmslow Road, Manchester M20 4BX, UK. wardcm@hotmail.com
Barrow
Katie M
KM
Stern
Peter L
PL
D000818:Animals; D015415:Biomarkers; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics*; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D019070:Cell Lineage / Q000187:drug effects / Q000235:genetics; D048429:Cell Size / Q000187:drug effects / Q000235:genetics; D002999:Clone Cells / Q000187:drug effects / Q000378:metabolism; D003470:Culture Media / Q000494:pharmacology*; D014644:Genetic Variation / Q000187:drug effects / Q000235:genetics*; D051379:Mice; D016900:Neurofilament Proteins / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects / Q000502:physiology; D014212:Tretinoin / Q000494:pharmacology
D015415:Biomarkers; D003470:Culture Media; D016900:Neurofilament Proteins; C066858:neurofilament protein NF 68; D014212:Tretinoin