J1細(xì)胞

數(shù)量： 大量

生長狀態(tài)： 貼壁生長

年限： embryo, blastocyst

品系： 129S4/SvJae

組織來源： inner cell mass

運(yùn)輸方式： 凍存運(yùn)輸

細(xì)胞形態(tài)： 其他

細(xì)胞類型： 胚胎干細(xì)胞

是否是腫瘤細(xì)胞： 0

物種來源： 小鼠

器官來源： 胚胎

ATCC Number： SCRC-1010?

Designations: J1

Depositors: R Jaenisch

J1細(xì)胞Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Mus musculus

Morphology: Spherical colony

Source: Organ: embryo

Strain: 129S4/SvJae

Tissue: inner cell mass

Cell Type: embryonic stem cell;

Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.

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.

Isolation: Isolation date: 1991

Applications: Published 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 J1細(xì)胞[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].

Cytogenetic Analysis: According to the depositor, this line has the following karyotype: normal 16/20 spreads; 4 show random artifacts.

Age: embryo, blastocyst

Gender: male

Comments: 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. The deposited line has been shown to be germline competent.

Propagation: ATCC complete growth medium: Mouse ES Cell Basal Medium

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Temperature: 37.0°C

Subculturing: Protocol: Establishing and maintaining your culture: To insure the highest level of viability, be sure to warm media to 37?C before using it on the cells.

Plate mitotically arrested MEF (CF-1) (ATCC SCRC-1040) as a feeder layer at approximately 1.5 to 2.0 X 106 cells/T25 at least one day before plating J1 cells (see product sheet for mitotically arrested MEF for protocol). One hour before thawing the vial of J1 ES cells, perform a 100% medium change using 10 ml of complete growth medium for ES cells.

J1細(xì)胞Thaw the vial 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 as soon as the contents are 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 xg for 5 min. Aspirate the supernatant and resuspend the pellet in 5 ml of complete growth medium for ES cells.

Add the 5 ml of cell suspension to the T75 flask containing feeder cells and 10 ml complete growth medium for ES cells.

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 to 2 days.

Subculturing Procedure: To insure the highest level of viability, be sure to warm media and Trypsin/EDTA to 37°C before using it on the 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 every 1 to 2 days is recommended. Plating densities should range from 3 to 4 X 106 cells/ T75. Note: If the colonies are close to or touching each other the culture is overgrown . Overgrowth will result in differentiation.

Prepare enough flasks with MEFs as stated above in step #1.

Aspirate the medium from the flask(s) with J1 ES cells.

Wash with PBS (Ca+2/Mg+2-free, ATCC cat# SCRR-2201).

Add 3.0 ml of 0.25% (w/v) Trypsin / 0.53 mM EDTA solution (ATCC cat # 30-2101) and place in incubator. After about one minute the ES colonies will dissociate and all cells will detach from the flask.

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.

Spin the cells at 270 xg for 5 min. Aspirate the supernatant.

Resuspend in 30 to 50 ml of fresh culture medium, depending on the split ratio.

Aspirate the medium from 4 to 7 feeder layer flasks and replace it with 15 ml/flask of J1 cell suspension.

Incubate the culture at 37°C in a humidified 5% CO2/95% air incubator. J1細(xì)胞Perform a 100% medium change every day, passage cells every 1 to 2 days.

Interval: Every one to two days

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:7 is recommended

Medium Renewal: Every day

Preservation: Freeze medium: Complete growth medium supplemented with an additional 10% FBS and 10% DMSO

Storage temperature: liquid nitrogen vapor phase

Related Products: Recommended medium: ATCC SCRR-2011

recommended serum: ATCC SCRR-30-2020

References: 89420: Tucker KL, et al. A transgenic mouse strain expressing four drug-selectable marker genes. Nucleic Acids Res. 25: 3745-3746, 1997. PubMed: 9278500

89599: 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

89600: Li E, et al. Targeted mutation of the DNA methyltransferase gene results in embryonic lethality. Cell 69: 915-926, 1992. PubMed: 1606615

16173447: 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

16173449: 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

16173450: 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

16173451: 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

16173452: 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

16173453: 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

16173454: 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

16173460: 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