運(yùn)輸方式： 凍存運(yùn)輸

ATCC Number： CRL-2110?

細(xì)胞類型： 其他細(xì)胞類型

是否是腫瘤細(xì)胞： 0

物種來(lái)源： 轉(zhuǎn)基因小鼠

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

器官來(lái)源： 肺

品系： FVB/N

細(xì)胞形態(tài)： 上皮樣

數(shù)量： 大量

年限： 5 month old

Designations: MLE 12

Depositors: JA Whitsett

Biosafety Level: 2 [CELLS CONTAIN PAPOVAVIRUS ]

MLE 12細(xì)胞Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Mus musculus, transgenic deposited as mouse, transgenic

Morphology: epithelial

Source: Organ: lung

Strain: FVB/N

Cell Type: epithelialSV40 transformed

Cellular Products: lung surfactant proteins B and C (SP-B, SP-C)

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.

Isolation: Isolation date: 1992

Applications: Lung surfactant proteins B and C were detected.

The cells secrete phospholipids in response to phorbol esters and ATP but not in response to forskolin.

This line was established in 1992 by Kathryn A. MLE 12細(xì)胞Wikenheiser from pulmonary tumors in a mouse transgenic for the SV40 large T antigen under the control of the promoter region of the human surfactant protein C gene.

Tumorigenic: Yes

Age: 5 month old

Gender: female

Comments: This line was established in 1992 by Kathryn A. Wikenheiser from pulmonary tumors in a mouse transgenic for the SV40 large T antigen under the control of the promoter region of the human surfactant protein C gene.

The cells express the mRNA for large T antigen.

Lung surfactant proteins B and C were detected.

The cells secrete phospholipids in response to phorbol esters and ATP but not in response to forskolin.

Propagation: ATCC complete growth medium: HITES medium supplemented with 2% fetal bovine serum

HITES medium with 2% fetal bovine serum is formulated at the ATCC as follows:

Dulbecco's medium : Ham's F12, 50:50 mix (ATCC 30-2006)

MLE 12細(xì)胞Insulin 0.005 mg/ml

Transferrin 0.01 mg/ml

Sodium selenite 30 nM

Hydrocortisone 10 nM

beta-estradiol 10 nM

HEPES 10 mM

L-glutamine 2 mM (in addition to that in the base medium)

fetal bovine serum 2%

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

Subculturing: 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 1:5 to 1:10 is recommended

Medium Renewal: Twice per week

Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO

Storage temperature: liquid nitrogen vapor phase

Doubling Time: 7 to 9 hrs

Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2006

References: 23101: Wikenheiser KA, et al. Simian virus 40 large T antigen directed by transcriptional elements of the human surfactant protein C gene produces pulmonary adenocarcinomas in transgenic mice. Cancer Res. 52: 5342-5352, 1992. PubMed: 1394139

23539: Wikenheiser KA, et al. Production of immortalized distal respiratory epithelial cell lines from surfactant protein C/simian virus 40 large tumor antigen transgenic mice. Proc. MLE 12細(xì)胞Natl. Acad. Sci. USA 90: 11029-11033, 1993. PubMed: 8248207