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產品名稱	SCC#10
商品貨號	B220605
Organism	Mus musculus, mouse
Tissue	inner cell mass
Cell Type	embryonic stem cell
Product Format	frozen
Morphology	Stem cell
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.] Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Age	embryo
Gender	Male
Strain	129X1/SvJ
Storage Conditions	liquid nitrogen vapor phase
Karyotype	40 XY, diploid
Derivation	The 129Sv/J ES line was derived from a 129Sv/J (129X1Sv/J in the current nomenclature) strain of mouse, allowing for production of knock-out and transgenic mice. 129Sv/J is a parental substrain of 129 strains and has a white-bellied, light chinchilla or albino coat color with pink eyed appearance.
Clinical Data	Male
Comments	This mouse ES cell line has been shown to be germline competent. 129Sv/J is a parental substrain of 129 strains and has a white-bellied, light chinchilla or albino coat color with pink eyed appearance. This strain is homozygous for Cdh23^ahl, the age-related hearing loss 1 mutation, resulting in progressive hearing loss with onset prior to 3 months of age (http://jaxmice.jax.org/strain/000691.html).
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	Subculturing Procedure 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 Daily maintain a sufficient number of flasks that have been pre-plated with MEFs in complete medium for feeder cells. 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 Aspirate the medium from the flask(s) containing ES cells. Wash with PBS Ca+2/Mg+2-free (ATCC SCRR-2201). 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. 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 x g for 5 min. Aspirate the supernatant. 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. 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. 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	Temperature: 37°C
Name of Depositor	TJ Ley
Deposited As	Mus musculus
Year of Origin	2003
References	Festing MF, et al. Revised nomenclature for strain 129 mice. Mamm. Genome. 10(8):836, 1999.[PubMed: 10430671] Gossler A, et al. Transgenesis by means of blastocyst-derived embryonic stem cell lines. Proc. Natl. Acad. Sci. USA 83: 9065-9069, 1986. PubMed: 3024164 Simpson EM, et al. Genetic variation among 129 substrains and its importance for targeted mutagenesis in mice. Nat. Genet. 16(1):19-27, 1997. PubMed: 9140391 Stevens LC. A new inbred subline of mice (129-terSv) with a high incidence of spontaneous congenital testicular teratomas. J. Natl. Cancer. Inst. 50(1): 235-242, 1973. PubMed: 4692863 Threadgill DW, et al. Genealogy of the 129 inbred strains: 129/SvJ is a contaminated inbred strain. Mamm. Genome. 8(6):390-393 1997. PubMed: 9166580 Jackson Laboratory Caputo JL. Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988 Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

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