General description

RESGRO Culture Medium is a specialty media that can be utilized to complement traditional murine Embryonic Stem (ES) cell culture media containing ESGRO^® murine LIF medium supplement. In contrast to routine ES cell culture using ESGRO supplemented medium, RESGRO Culture Medium is recommended for a number of specialized applications.

Application

Rescue of established ES cell lines

RESGRO Culture Medium has the capacity to rescue established ES cell lines that have started drifting, and either generate low percentage chimeras or have lost germline transmission capability. Differentiation, which is present in the ES cells but not visible with traditional medium, will become recognizable when using RESGRO Culture Medium. After 2 passages, a clear difference is seen between differentiated and undifferentiated ES cells, at which time undifferentiated cells can be selected for by sub-cloning.

Murine ES cell derivation

The efficiency of ES cell derivation is greatly strain dependent. To date, very few murine ES cell lines are available from inbred strains other than 129 strains, and those derived 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, these cell lines generate chimeras less efficiently and contribute less frequently to the germline.

RESGRO Culture Medium enables the efficient derivation and maintenance of ES cell lines from several inbred mouse strains, including certain strains that were previously considered to be non-permissive for ES cell derivation. A recent study demonstrated that RESGRO allowed the derivation of ES cell lines from inbred strains other than 129 (including FVB, a strain previously considered to be non-permissive for ES cell derivation and C57Bl/6N, BALB/c, 129/SvEv and DBA/2N mouse strains)1.

Culture of ES cells without a fibroblast feeder layer

RESGRO Culture Medium allows for the culture of ES cells on bare (gelatinized) culture dishes. Even in the absence of a fibroblast feeder layer ES cells maintain their undifferentiated character and their germline transmission capability for at least 5 passages, when cultured with RESGRO Culture Medium. After trypsinization pure ES cell suspensions without fibroblast cells can be obtained. Fibroblast cells will no longer interfere during blastocyst injections, diploid aggregations, tetraploid aggregations and electroporations.

The RESGRO Culture Medium is a specialty media that can be utilized to complement traditional murine Embryonic Stem (ES) cell culture media containing ESGRO murine LIF medium supplement.

Storage and Stability

Upon receipt, RESGRO Culture Medium should be stored at -80°C until required for use. When stored properly, product is stable until the expiration date stated on the label. Once opened, the unused portion can be stored at 4°C for at least 4 weeks. Protect from light during storage. Avoid repeated freeze/thaw cycles.

It is recommended that the product be discarded following expiry. Be aware that the addition of certain supplements can effect product stability and characteristics.

Note: Turbidity and flocculent material may be present after thawing or after prolonged freezer and/or refrigerated storage.

We recommend that in case of flocculent material or turbidity, to use a GP Express Plus Membrane (Stericup from Millipore - ref. SCGPU05RE) in combination with a standard 70 mm fiberglass prefilter for 500 mL bottle top filters.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

ESGRO is a registered trademark of Merck KGaA, Darmstadt, Germany

PLURISTEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

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