LLC-PK1A細(xì)胞

ATCC Number： CL-101.1?

相關(guān)**： 正常

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

數(shù)量： 大量

運(yùn)輸方式： 凍存運(yùn)輸

器官來(lái)源： 腎臟

是否是腫瘤細(xì)胞： 0

物種來(lái)源： 豬

Designations: LLC-PK1A

Depositors: Eli Lilly & Co.

LLC-PK1A細(xì)胞Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Sus scrofa

Morphology:

Source: Organ: kidney

Disease: normal

Cellular Products: plasminogen activator

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. LLC-PK1A細(xì)胞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.

Propagation: ATCC complete growth medium: The base medium for this cell line is Medium 199 with Earle's BSS with 1.5 g/L to 2.2 g/L sodium bicarbonateTo make the complete growth medium, add the following components to the base medium:

5% fetal bovine serum

Temperature: 37.0°C

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

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

Medium Renewal: Twice per week

Protocol: Volumes used in this protocol are for 75 sq cm flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

Remove and discard culture medium.

LLC-PK1A細(xì)胞Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which 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 37C to facilitate dispersal.

Add 2.0 to 3.0 ml of complete growth medium and aspirate cells by gently pipetting

Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.

Incubate cultures at 37C.

Preservation: culture medium, 90%; additional serum, 5%; DMSO, 5%

References: 3520: Hull RN, Huseby RM. LLC-PK1A細(xì)胞Enhanced production of plasminogen activator. US Patent 3,904,480 dated Sep 9 1975