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> Embryonic Stem Cells
Embryonic Stem Cell Medium
(Reference #52-TX-WES) ES culture medium
To maintain ES cells in a pluripotent state, it is important to refeed the ES cells daily with fresh stem cell medium. Traditional stem cell culture medium is a mix of DMEM, FBS and some additional compounds like LIF and other growing factors, non-essential amino acids, sodium pyruvate, glutamine, ß-mercapto-ethanol, Penicillin/Streptomycin,ˇ
Unique characteristic of the TX-WES cell culture medium:
The TX-WES medium is a LIF-free medium, which allows the derivation of ES cell lines from inbred strains other than 129 and the rescue of traditional ES cell lines that started drifting.
The efficiency of ES cell derivation is greatly strain dependent. Very few ES cell lines are currently available from inbred strains (mostly C57Bl/6) other than the 129 strains, and those have generally been obtained with low success rates. Furthermore, ES cells derived from other strains than 129 are in general more difficult to propagate in vitro. Especially at high passage number and after genetic manipulation, they can generate chimeras less efficiently, and they contribute less frequently to the germline. The TX-WES medium enables the efficient derivation and maintenance in culture of ES cell lines in several inbred mouse strains, including certain strains that were previously considered to be non-permissive for ES cell derivation.
In addition, the TX-WES cell medium has the capacity to rescue traditional ES cell lines that started drifting and only generate low percentage chimeras or have lost germ line transmission capability. Differentiation which is present in the cells, but not visible with traditional medium, will become recognisable with the TX-WES cell medium: after 2 passages, a clear difference is seen between differentiated and undifferentiated ES cells, and at that moment, a subcloning can be performed according to the Thromb-X procedure to pick out the undifferentiated cells.
Other characteristics of the TX-WES cell culture medium:
• The TX-WES medium is 'ready-to-use' . Only add sterile L-glutamine (200mM; 10ml to 500ml TX-WES) and start working.
• There is no need to test the medium before use. All compounds and the complete medium are tested in advance by our company.
Tests performed are:
Mycoplasm and other pathogens
Tested for its capacity to culture ES cells
Tested for its capacity to allow ES cell derivation from inbred FVB mice
• The medium can be stored at ?C80ˇăC for at least 2 years
• If needed, only use cellulose acetate filters
• The TX-WES medium is available in bottles of 250ml or 500ml.
• We do not advise repeated thawing and freezing of the medium.
(Reference #51-C57B1/6N) • All MEF cell strains are available at passage number 3 (or at a lower passage number upon request).
• The cells are MAP tested (free for mycoplasma and other pathogens) and have been tested for their capacity to sustain ES cell growth with germline transmission capability.
• MEF cells are available per unit: each unit contains two cryogenic vials with about 3x106 cells per vial.
• The MEF cells are not yet inactivated and expansion is still possible. However, we do not advise to use them at a passage number higher as 5
Neo+/Neo+ MEF cells
Hprt- MEF cells