L Cells細(xì)胞

品系： C3H/An

年限： 100 days

組織來源： subcutaneous connective tissue; areolar and adipose

ATCC Number： CRL-2648?

細(xì)胞類型： 其他細(xì)胞類型

生長狀態(tài)： 貼壁生長

數(shù)量： 大量

是否是腫瘤細(xì)胞： 0

物種來源： 小鼠

細(xì)胞形態(tài)： 成纖維樣

L Cells細(xì)胞運(yùn)輸方式： 凍存運(yùn)輸

Designations: L Cells

Depositors: R Nusse

Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Mus musculus deposited as mouse

Morphology: fibroblast

Source: Tissue: subcutaneous connective tissue; areolar and adipose

Strain: C3H/An

Cell Type: fibroblast fibroblast;

Permits/Forms: L Cells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 (Roche Transfection Reagents)

Age: 100 days

Gender: male

Comments: It is the parental line for the L Wnt-3A cell line (ATCC CRL-2647) and the L Wnt-5A cell line (ATCC CRL-2814). It is used to obtain control conditioned medium for comparison to Wnt-3A or Wnt-5A conditioned medium from L Wnt-3A cells or L Wnt-5A cells.

The cell line named L Cells is closely related or identical to L-M(TK-) cells (ATCC CCL-1.3).

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%.

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

Temperature: L Cells細(xì)胞37.0°C

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.

Protocol for control conditioned medium:

Split the cells 1:10 in 10 ml culture medium in 10 cm petri dishes and let the cells grow for 4 days (approximately to confluency).

Take off the medium and sterile filter. This is the first batch of medium.

Add 10 ml fresh culture medium and culture for another 3 days.

Take off the medium and sterile filter. This is the second batch of medium. Discard the cells, because they will be overgrown.

Mix the first batch and second batch of medium 1:1. This is the L Cell conditioned medium. It is stable at 4°C and can be frozen.

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:10 is recommended

Medium Renewal: Every 2 to 3 days

Preservation: Freeze medium: L Cells細(xì)胞Complete growth medium supplemented with 5% (v/v) DMSO

Storage temperature: liquid nitrogen vapor phase

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

recommended serum:ATCC 30-2020

derivative:ATCC CRL-2647

derivative:ATCC CRL-2814

References: 90267: Willert K, et al. Wnt proteins are lipid-modified and can act as stem cell growth factors. Nature 423: 448-452, 2003. PubMed: 12717451