L-M(TK-) [LM(tk-), LMTK-]細(xì)胞

是否是腫瘤細(xì)胞： 0

物種來源： 小鼠

品系： C3H/An

組織來源： subcutaneous connective tissue; areolar and adipose

生長狀態(tài)： 貼壁生長

ATCC Number： CCL-1.3?

運(yùn)輸方式： 凍存運(yùn)輸

數(shù)量： 大量

年限： 100 days

細(xì)胞形態(tài)： 成纖維樣

L-M(TK-) [LM(tk-), LMTK-]細(xì)胞Designations: L-M(TK-) [LM(tk-), LMTK-]

Depositors: TR Chen

Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Mus musculus

Morphology: fibroblast

Source: Tissue: subcutaneous connective tissue; areolar and adipose

Strain: C3H/An

Permits/Forms: L-M(TK-) [LM(tk-), LMTK-]細(xì)胞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.

Applications: transfection host

Tumorigenic: Yes

Antigen Expression: H-2k

Cytogenetic Analysis: modal number = 45; range = 42 to 48.

The stemline chromosome number is hyperdiploid with the 2S component occurring at 3.8%. Twenty-one markers were common to nearly all S metaphases studied. Of these, 13 were two-armed chromosomes, and eight, acrocentric. Four markers were isochromosomes by Robertsonian translocations from chromosomes 4, 12, 14 and 15. Normal X, 12, 14, 15 and 16 were absent. The X chromosome was present as an arm of Rb(X.3). (See Cancer Genet. Cytogenet. 24: 95-107, 1987 for a detailed karyotype description.)

Age: 100 days

Gender: male

Comments: The cells are 5-bromo-2'-deoxyuridine resistant (BUDR, BrdUrd) and thymidine kinase (TK) deficient.

HAT sensitive.

Tested and found negative for ectromelia virus (mousepox).

Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.

Temperature: 37.0°C

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

Subculturing: L-M(TK-) [LM(tk-), LMTK-]細(xì)胞Protocol:

Remove and discard culture medium.

Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.

Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).

Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37?C to facilitate dispersal.

Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.

Add appropriate aliquots of the cell suspension to new culture vessels.

Incubate cultures at 37?C.

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended

Medium Renewal: Twice per week

Preservation: Freeze medium: complete growth medium, 95%; DMSO, 5%

Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002

recommended serum:ATCC 30-2020

parental cell line:ATCC CCL-1

References: 868: Kit S, et al. L-M(TK-) [LM(tk-), LMTK-]細(xì)胞Deletion of thymidine kinase activity from L cells resistant to bromodeoxyuridine. Exp. Cell Res. 31: 297-312, 1963. PubMed: 14056989

21410: Chen TR. Cytogenetics of somatic cell hybrids. II. Modal karyotypes and mosaicism in mouse mutant cell lines: LM(tk-), A9, and Rag. Cancer Genet. Cytogenet. 24: 95-107, 1987. PubMed: 3466673

25904: Hsu TC, Somers CE. Properties of L cells resistant to 5-bromodeoxyuridine. Exp. Cell Res. 26: 404-410, 1962. PubMed: 14449548

25907: Boone C, et al. Assignment of three human genes to chromosomes (LDH-A to 11, TK to 17, and IDH to 20) and evidence for translocation between human and mouse chromosomes in somatic cell hybrids (thymidine kinase-lactate dehydrogenase A-isocitrate dehydrogenase-C-11, E-17, and F-20 chromosomes). Proc. Natl. Acad. Sci. USA 69: 510-514, 1972. PubMed: 4110482

25910: Peterson JL, McBride OW. Cotransfer of linked eukaryotic genes and efficient transfer of hypoxanthine phosphoribosyltransferase by DNA-mediated gene transfer. Proc. Natl. Acad. Sci. USA 77: 1583-1587, 1980. PubMed: 6929511