ESGRO Complete PLUS Clonal Grade Medium, 100ml

QC PROTOCOL:

Low density plating assay:
1. Plate feeder independent early passage ES cells onto gelatin coated 10 cm plastic tissue culture dish at 1000 cells/10 cm dish in 10-20 mL of Clonal Grade Medium.
2. Change medium every 2-4 days.
3. Monitor for clonal growth with alkaline phosphatase stain.

Colony formation should occur over 4-6 days. The majority of colonies should be alkaline phosphatase positive and show no signs of differentiation.

Suitable for ES cell derivation, transfection and selection, in addition to general cell passaging and growth.
Media can be used on feeder dependent/independent ES cell lines.

Important passaging information
Use of one of the below listed cell dissociation solutions is imperative while passaging ES cells using ESGRO Complete products. Standard trypsin protocols used to passage ES cells grown in serum containing media typically result in over trypsinization causing the cells to lift off the plates and form abnormal ball-like structures.
Recommended products include:
ESGRO Complete^™ Accutase (Cat # SF006)
ESGRO Complete Trypsin Solution (proprietary) (Cat # SF007)
ESGRO Complete Enzyme-Free Dissociation Solution (proprietary) (Cat # SF009)

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.

ESGRO Complete PLUS Clonal Grade Medium is a defined serum-free medium provided with a selective GSK3β inhibitor to enhance viability of mouse embryonic stem (ES) cells and increased maintenance of pluripotency in the absence of serum and feeder cells. This medium is intended for mouse ES cell expansion and growth at clonal density in serum-free conditions. The mouse ES cells should maintain germ line competency.

Replaces: SF001-100

• Upon receipt, store clonal grade medium and supplement in the dark at -20°C. Refer to lot expiration date on labels.
• Thawed clonal grade medium should be stored in the dark at 4°C for one month. Do not refreeze.
• Supplement may be thawed at room temperature, aliquoted and stored in the dark at 4°C for two months.
• After supplement is added to clonal grade medium, store in the dark at 4°C for up to
two weeks. Do not refreeze.

1. ESGRO Complete Clonal Grade Medium: (Catalogue No. SF001-C) One (1) 100 mL bottle containing original clonal grade medium. Complete formulation containing self-renewal factors, no need to add LIF. pH 7.1-7.3

2. GSK3β Inhibitor Supplement: (Catalogue No. SF012-050) One (1) 50 µL vial containing selective GSK3β inhibitor.

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

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

cOmplete is a trademark of Roche