MC2/3細(xì)胞

ATCC Number： CRL-2143?

運(yùn)輸方式： 凍存運(yùn)輸

生長狀態(tài)： 貼壁生長

細(xì)胞形態(tài)： 成纖維樣

數(shù)量： 大量

細(xì)胞類型： 其他細(xì)胞類型

是否是腫瘤細(xì)胞： 0

物種來源： 倉鼠

Designations: MC2/3

Depositors: RL Stallings

Biosafety Level: 1

MC2/3細(xì)胞Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Cricetulus griseus

Morphology: fibroblast

Source: Cell Type: somatic cell hybrid;

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Comments: MC2/3 is a mouse chromosome 8 specific monochromosome hybrid cell line.

This hybrid resulted from the fusion of adenine phosphoribosyl transferase negative (APRT -) Chinese hamster ovary cells with micronuclei from near euploid thymidine kinase negative (TK -) CAK mouse cells.

Hybrids containing mouse chromosome 8 (which has the APRT locus) were selected in medium containing 0.075 mM adenine, 800 nM aminopterin and 0.016 mM thymidine (AAT).

MC2/3細(xì)胞The mouse chromosome 8 could be from either the C57BL/6J or ARK/J inbred strain, since CAK cells were derived from an F1 hybrid of those two strains.

Mouse chromosome 8 may be isolated with 96% purity.

The depositor has reported the presence of one copy of mouse chromosome 8 in each Chinese hamster cell by G band cytogenetics, fluorescence in situ hybridization (FISH) and PCR analysis.

However, FISH analysis of ATCC frozen stocks by the depositor using biotinylated total genomic mouse DNA as probe indicates that 7 of 9 metaphases examined had a single mouse acrocentric chromosome present in a hamster background.

Two of nine metaphases showed different rearrangements of the mouse acrocentric.

An examination of interphase nuclei indicates that about 18% of the cells are tetraploid showing two distinct hybridization signals from the mouse while 82% are near diploid exhibiting one distinct signal.

The level of tetraploidy may increase with additional passage.

Propagation: ATCC complete growth medium: Dulbecco's modified Eagle's medium containing 0.075 mM adenine, 800 nM aminopterin, and 0.016 mM thymidine, 80%; fetal bovine serum, 20%

Subculturing: Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended

Medium Renewal: MC2/3細(xì)胞Twice per week

Remove spent medium, add fresh 0.25% trypsin, 0.03% EDTA solution, rinse and remove trypsin. Let the culture sit at room temperature (or at 37C) until the cells detach. Add fresh medium, aspirate and dispense into new flasks.

Preservation: Culture medium, 95%; DMSO, 5%

References: 22418: Stallings RL, Deaven LL. A mouse monochromosome 8 somatic cell hybrid: a reagent for chromosome 8 isolation. Mamm. Genome 5: 572-573, 1994. PubMed: 8000142