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embryo, blastocyst
Designations: | B6/BLU | ||
Depositors: | TJ Ley | ||
Biosafety Level: | 1 | ||
Shipped: | frozen | ||
Medium & Serum: | See Propagation | ||
Organism: | Mus musculus | ||
Morphology: | stem cell |
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Source: | Strain: C57BL/6 Organ: embryo Tissue: inner cell mass Cell Type: stem cell embryonic stem cell; |
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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: 1998 | ||
Applications: | The B6/BLU ES cell line was derived from a C57BL/6 transgenic line that contains a LacZ reporter. | ||
Cytogenetic Analysis: | 40 XY, diploid | ||
Age: | embryo, blastocyst | ||
Gender: | male | ||
Comments: | This mouse ES cell line has been shown to be germline competent. The B6/BLU ES cell line was derived from a C57BL/6 transgenic line that contains a LacZ reporter. The transgene is a -globin LacZ fusion expressed exclusively in peripheral red blood cells. The transgene was assembled in pUC19, and contains the 1.9 kb KpnI-PvuII human 54 HS-2 fragment, upstream from the marked �-globin transgene. The 5' part of the human �-globin gene was fused to a 3' kb NcoI-BglII fragment obtained from pLacD. The 3' part of the -globin gene consisted of a 2.8 kb BamHI-XbaI fragment that contains the 3' end of exon 2, intron 2, exon 3 and 3' flanking sequence. 5' HS-2 was inserted in the genomic (5' to 3') orientation with respect to the transgene. The transgene was isolated from the plasmid vector backbone by cleavage with XhoI and SalI .Instead of determining chimerism visually by coat color or assessing chimerism genotypically in a tail DNA, chimerism is assessed quantitatively in the mesoderm by assessing the LacZ expression in a single drop of tail blood obtained at weaning. [16172710] | ||
Propagation: | ATCC complete growth medium: Mouse ES Cell Basal Medium Temperature: 37.0°C Growth conditions: Use a feeder layer, LIF and frequent subcultures to prolong the undifferentiated state Atmosphere: air, 95%; carbon dioxide (CO2), 5% |
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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.
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.
Interval: every 2 to 3 days Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:7 is recommended Medium Renewal: Every day. |
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Preservation: | Storage temperature: liquid nitrogen vapor phase Freeze medium: Complete growth medium supplemented with an additional 10% FBS and 10% DMSO |
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References: | 16172708: Hughes, DE et al. Genetic variation in C57BL/6 ES cell line. Mamm. Genome 18: 549-558, 2007. PubMed: 17828574 16172709: Ware, BC, et al. Utiltiy of a C57BL/6 ES line versus 129 ES lines for targeted mutations in mice. Transgenic Res. 12: 743-746, 2003. PubMed: 14713204 16172710: Graubert A T, et al. Stochastic, stage-specific mechanisms account for the variegation of a human globin transgene. Nucleic Acids Research. 26 (12) :2849-2858, 1998. PubMed: 9611227 |
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