C57BL/6 (RMCL-00260301)
Cat. No.: RMCL-00260301
Category: Cell Lines
INQUIRY
1 vial
C57BL/6 is a spherical colony that was isolated from a mouseembryo. This cell in coat-color chimera production was high using c2J blastocysts while FVB blastocysts produced a low number of chimeras. The clonal embryonic stem cell line ES C57BL/6 was derived from a strain C57BL/6J (B6) mouse blastocyst.
Product Features
| BSL | BSL-1 |
|---|---|
| Product Format | Frozen |
| Applications | Stem cell research |
| Specific Applications | Coat-color chimera production was high using c2J blastocysts while FVB blastocysts produced a low number of chimeras.The clonal embryonic stem cell line ES C57BL/6 was derived from a strain C57BL/6J(B6) mouse blastocyst.The ES cells were shown to populate the germ line of two host blastocyst donors, FVB/NJ (FVB) and the coisogenic strain C57BL/6-Tyrc-2J(c2J). |
| Product Type | Animal cells |
| Strain | C57BL/6 |
| Organism | Mus musculus, mouse |
| Tissue | Embryo |
| Morphology | Spherical colony |
| Cell Type | Embryonic Stem Cell |
QC
| Mycoplasma Contamination | Not detected |
|---|
Doner Information
| Age | Embryo |
|---|
Storage & Shipping
| Storage Conditions | Vapor phase of liquid nitrogen |
|---|---|
| Unpacking and Storage Instructions | Check all containers for leakage or breakage. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below -130°C, preferably in liquid nitrogen vapor, until ready for use. |
| Growth Properties | Adherent |
| Temperature | 37°C |
| Atmosphere | 95% Air, 5% CO2 |
| Handling procedure | Complete Medium for Feeder Cells Feeder cells may be grown in medium containing fewer growth factors than those required by the ES cells. Consult the product sheet provided for the feeder cells you wish to use for medium requirements. Feeder cells should be initiated 24-48 hours prior to inoculating with embryonic stem (ES) cells. Feeder Cells recommends culturing ES-C57BL/6 on mouse embryonic fibroblasts (MEFs) that have been mitotically arrested by either irradiation or treatment with Mitomycin-C. ES-C57BL/6 cells have been cultured on mitotically arrested MEF (CF-1). At least one day before plating the ES cells, prepare the desired combination of flasks with feeder cells to accommodate an initial ES cell seeding density of 30,000 cells/cm2 to 50,000 cells/cm2. Refer to the batch specific information provided on the last page of the product information sheet for the total number of viable cells recovered from this lot of SCRC-1002. Plate mitotically arrested mouse embryonic fibroblasts (MEFs) as a feeder layer at approximately 55,000 feeder cells/cm2 in complete medium for feeder cells. Refer to the product sheet for mitotically arrested MEF for detailed handling instructions. Feeder cells should be used within one week of plating. It is best to use feeder cells within 24 to 48 hours of initiation. Embryonic Stem (ES) Cells 30 Minutes Prior to Handling Cells – Pre-warm complete growth medium for ES cells at 37°C for at least 30 minutes before adding to cells. One Hour Prior to Thawing the ES Cells – Perform a 100% medium change for the MEFs using complete growth medium for ES cells. Thaw the vial of ES cells by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 90 seconds). Remove the vial from the water bath before the contents are completely thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions. Transfer the vial’s contents plus 5 mL of complete growth medium for ES cells to a 15 mL centrifuge tube. Use an additional 1 mL of media to rinse the vial and transfer the liquid to the 15 mL tube. Add 4 ml of complete growth medium for ES cells to bring the total volume to 10 mL. Spin the cells at 270 x g for 5 min. Aspirate the supernatant and resuspend the pellet in 2 mL of complete growth medium for ES cells. Add the 2 mL of cell suspension to the appropriate size flask containing feeder cells and fresh complete growth medium for ES cells (see batch specific information). ES cells should be plated at a density of 30,000 – 50,000 cells/cm2. Incubate the culture at 37°C in a humidified 5% CO2/95% air incubator. Routine Handling Perform a 100% medium change every day. Passage the cells every 1 to 2 days. If the colonies are close to or touching each other the culture is overgrown. Overgrowth will result in differentiation. Make sure that you have prepared a sufficient number of flasks pre-plated with MEF feeder layers to support frequent passage of the ES cells. |
For research use only. Not for clinical use.
