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ATCCSCRC-1020 SCC#10
基本信息
资源编号 ATCCSCRC-1020
资源名称 SCC#10
种属 Mus musculus, mouse inner cell mass
类型 embryonic stem cell
形态 Stem cell
提供形式 frozen
安全等级 1
培养方法
培养基 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.
传代方法 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.
生长条件 Temperature: 37°C
Growth condition: feeder cells required.
存储条件 liquid nitrogen vapor phase;Liquid nitrogen vapor phase
详细说明
描述 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 Cdh23ahl, 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).
Age embryo
Gender Male
Strain 129X1/SvJ
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
Year of Origin 2003
Restrictions Prior to purchase, for-profit commercial institutions must obtain a license agreement. For instructions on how to proceed, please contact Washington University in St. Louis' Office of Technology Management Development at kratochj@wustl.edu or 314-747-0923.
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