MEF (DR4)
SCRC-1045 ™
SCRC-1045 ™
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ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.
Can be used to produce feeder cells with multiple drug resistance.
SCRC-1045 cells can be used as a feeder layer to support the growth of engineered embryonic stem (ES) cells with multiple drug selections and for the maintenance of ES cells in the undifferentiated state.
Dr. Rudolf Jaenisch at the Massachusetts Institute of Technology developed the DR4 mouse strain by the intercrossing of three different strains: one bearing resistance genes neoR and puroR, a second bearing the resistance gene hygR, and a third bearing a natural deletion encompassing the Hprt gene. A series of matings incorporated all 4 drug resistance genes into the strain (Tucker et al., 1997; PubMed: 9278500). The background of the original DR4 strain was a mix of 129S4/SvJae, 129P2/OlaHsd, BALB/c, and C57BL/6.
It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
Note: Volumes used in this protocol are for 75cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual Of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:7 is recommended
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Tucker KL, et al. A transgenic mouse strain expressing four drug-selectable marker genes. Nucleic Acids Res. 25: 3745-3746, 1997. PubMed: 9278500
Nagy A, et al. Manipulating The Mouse Embryo: A Laboratory Manual. Third Edition: Cold Spring Harbor Press; 2003.
Freshney RI. Culture of Animal Cells: A Manual of Basic Technique, 5th edition. New York: Wiley Liss; 2005. For more information on enzymatic dissociation and subculturing of cell lines see Chapter 13.