J1 (ATCC® SCRC-1010)

Organism: Mus musculus, mouse  /  Cell Type: embryonic stem cell  /  Tissue: inner cell mass  / 

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Organism Mus musculus, mouse
Tissue inner cell mass
Cell Type embryonic stem cell
Product Format frozen
Morphology Spherical colony
Culture Properties Adherent
Biosafety Level 1

Appropriate safety procedures should always be used with this material. Laboratory safety is discussed in the following publication: Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS Publication No. (CDC) 93-8395. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office (2007). The entire text is available online at

http://www.cdc.gov/biosafety/publications/bmbl5/index.htm.

Age embryo, blastocyst
Gender Male
Strain 129S4/SvJae
Applications J1 cells have been successfully employed in several different applications, including, but not limited to:
  • Biomarkers: Included in microarray studies aimed at developing methods for detecting novel marker ES genes [PubMed: 17875203], as well as biomarker signature patterns expressed during differentiation [PubMed: 17605829]. 
  • Embryonic development: Used to study X chromosome inactivation [PubMed: 11713286 and PubMed:9618446] and DNA methylation involved in gene regulation [PubMed: 17182866,PubMed:12370304, PubMed: 8917520 and PubMed: 11884600]. 
  • Cancer Research: Cre-Lox generation of a conditional VHL null allele in J1 cells has been used to generate Chimeric mice for the study von Hippel-Lindau (VHL) syndrome [PubMed: 11171994].
Storage Conditions liquid nitrogen vapor phase
Karyotype According to the depositor, this line has the following karyotype: normal 16/20 spreads; 4 show random artifacts.
Derivation
The J1 ES line was derived from a male agouti 129S4/SvJae embryo [PubMed: 1606615]. The J1 cells deposited at the ATCC were recloned from Dr. En Li's P6 J1 ES cells. 
Clinical Data
male
The J1 ES line was derived from a male agouti 129S4/SvJae embryo [PubMed: 1606615].
Comments
The deposited line has been shown to be germline competent.
Complete Growth Medium Grow ES cells in Mouse ES Cell Basal Medium (ATCC SCRR-2011) that has been supplemented with the following components:
1. 0.1 mM 2-mercaptoethanol (Life Technologies Cat. No. 21985-023)
2. 1,000 U/mL mouse leukemia inhibitory factor (LIF) (Millipore Cat. No. ESG1107)
3. 10% to 15% ES-Cell Qualified FBS (ATCC® SCRR-30-2020) or an ES cell qualified serum replacement
Complete Growth Medium for Mouse ES Cells is stable for 14 days when stored at 2°C to 8°C.
Subculturing

Note: To insure the highest level of viability, pre-warm media and Trypsin/EDTA to 37ºC before adding to cells. Volumes used in this protocol are for T75 flasks. Proportionally adjust the volumes for culture vessels of other sizes. A split ratio of 1:4 to 1:7 is recommended.

Feeder Cell Preparation for Subcultures

  1. Daily maintain a sufficient number of flasks that have been pre-plated with MEFs in complete medium for feeder cells.
  2. One hour before subculturing the ES cells, perform a 100% medium change for the MEFs using complete growth medium for ES cells.

Dissociation and Transfer of ES Cells

  1. Aspirate the medium from the flask(s) containing ES cells.
  2. Wash with PBS Ca+2/Mg+2-free (ATCC® SCRR-2201).
  3. Add 3.0 mL of 0.25% (w/v) Trypsin / 0.53 mM EDTA solution (ATCC® 30-2101) and place in incubator. After about one minute the ES colonies will dissociate and all cells will detach from the flask.
  4. Dislodge the cells by gently tapping the side of the flask then wash the cells off with 7-10 mL of fresh culture medium. Triturate cells several times with a 10 mL pipette in order to dissociate the cells into a single-cell suspension.
  5. Spin the cells at 270 x g for 5 min. Aspirate the supernatant.
  6. Resuspend in enough complete growth medium for ES cells to reseed new vessels at the desired split ratio (i.e. a split ratio of 1:4 to 1:7 is recommended). Perform a cell count to determine the total number of cells. ES cells should be plated at a density of 30,000 – 50,000 cells/ cm2.
  7. Add separate aliquots of the cell suspension to the appropriate size flask containing feeder cells and add an appropriate volume of fresh complete growth medium for ES cells to each vessel.
  8. Incubate the culture at 37°C in a humidified 5% CO2/95% air incubator. Perform a 100% medium change every day, passage cells every 1-2 days.
Cryopreservation
Liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Name of Depositor R Jaenisch
Year of Origin 1991
References

Stevens LC. A new inbred subline of mice (129-terSv) with a high incidence of spontaneous congenital testicular teratomas. J. Natl. Cancer Inst. 50: 235-242, 1973. PubMed: 4692863

Li E, et al. Targeted mutation of the DNA methyltransferase gene results in embryonic lethality. Cell 69: 915-926, 1992. PubMed: 1606615

Krzyzanowski PM, et al. Identification of novel stem cell markers using gap analysis of gene expression data. Genome Biology 8(9): R193, 2007. PubMed: 17875203

Luikenhuis S, et. al. Antisense transcription through the Xist locus mediates Tsix function in embryonic stem cells. Mol. Cell Biol. 21(24): 8512?8520, 2001. PubMed: 11713286

Goto T, et al. Regulation of X-Chromosome inactivation in development in mice and humans. Micro. Molec. Biol. Rev. (ASM) 62(2): 362-378, 1998. PubMed: 9618446

Yap DYL, et al. Using biomarker signature patterns for an mRNA molecular diagnostic of mouse embryonic stem cell differentiation state. BMC Genomics 8: 210, 2007. PubMed: 17605829

Haase VH, et al. Vascular tumors in livers with targeted inactivation of the von Hippel?Lindau tumor suppressor. Proc. Natl. Acad. Sci. 98(4): 1583?1588, 2001. PubMed: 11171994

Oda M, et al. DNA methylation regulates long-range gene silencing of an X-linked homeobox gene cluster in a lineage-specific manner. Genes Dev. 20(24): 3382?3394, 2006. PubMed: 17182866

Lorincz MC, et al. DNA methylation density influences the stability of an epigenetic imprint and Dnmt3a/b-independent de novo methylation. Mol. Cell Biol. 22(21): 7572?7580, 2002.PubMed: 12370304

Tucker KL, et al. A transgenic mouse strain expressing four drug-selectable marker genes. Nucleic Acids Res. 25: 3745-3746, 1997. PubMed: 9278500

Biniszkiewicz D, et al. Dnmt1 overexpression causes genomic hypermethylation, loss of imprinting, and embryonic lethality. Mol. Cell Biol. 22(7): 2124?2135, 2002. PubMed: 11884600

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Restrictions

Prior to purchase, for-profit commercial institutions must obtain a license agreement. For instructions on how to proceed, please contact ATCC's Office of Licensing and Business Development at licensing@atcc.org or 703 365 2773.

References

Stevens LC. A new inbred subline of mice (129-terSv) with a high incidence of spontaneous congenital testicular teratomas. J. Natl. Cancer Inst. 50: 235-242, 1973. PubMed: 4692863

Li E, et al. Targeted mutation of the DNA methyltransferase gene results in embryonic lethality. Cell 69: 915-926, 1992. PubMed: 1606615

Krzyzanowski PM, et al. Identification of novel stem cell markers using gap analysis of gene expression data. Genome Biology 8(9): R193, 2007. PubMed: 17875203

Luikenhuis S, et. al. Antisense transcription through the Xist locus mediates Tsix function in embryonic stem cells. Mol. Cell Biol. 21(24): 8512?8520, 2001. PubMed: 11713286

Goto T, et al. Regulation of X-Chromosome inactivation in development in mice and humans. Micro. Molec. Biol. Rev. (ASM) 62(2): 362-378, 1998. PubMed: 9618446

Yap DYL, et al. Using biomarker signature patterns for an mRNA molecular diagnostic of mouse embryonic stem cell differentiation state. BMC Genomics 8: 210, 2007. PubMed: 17605829

Haase VH, et al. Vascular tumors in livers with targeted inactivation of the von Hippel?Lindau tumor suppressor. Proc. Natl. Acad. Sci. 98(4): 1583?1588, 2001. PubMed: 11171994

Oda M, et al. DNA methylation regulates long-range gene silencing of an X-linked homeobox gene cluster in a lineage-specific manner. Genes Dev. 20(24): 3382?3394, 2006. PubMed: 17182866

Lorincz MC, et al. DNA methylation density influences the stability of an epigenetic imprint and Dnmt3a/b-independent de novo methylation. Mol. Cell Biol. 22(21): 7572?7580, 2002.PubMed: 12370304

Tucker KL, et al. A transgenic mouse strain expressing four drug-selectable marker genes. Nucleic Acids Res. 25: 3745-3746, 1997. PubMed: 9278500

Biniszkiewicz D, et al. Dnmt1 overexpression causes genomic hypermethylation, loss of imprinting, and embryonic lethality. Mol. Cell Biol. 22(7): 2124?2135, 2002. PubMed: 11884600