CRL-1740 LNCaP clone FGC 人前列腺癌細(xì)胞
ATCC® Number: CRL-1740?
Designations: LNCaP clone FGC
Depositors: JS Horoszewicz
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent, single cells and loosely attached clusters
Organism: Homo sapiens (human)
Morphology: epithelial
Source: Organ: prostate
Disease: carcinoma
Derived from metastatic site: left supraclavicular lymph node
Cellular Products: human prostatic acid phosphatase; prostate specific antigen [21889]
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Isolation: Isolation date: 1977
Applications: transfection host (Nucleofection technology from Lonza
Roche FuGENE® Transfection Reagents)
Receptors: androgen receptor, positive; estrogen receptor, positive [23045]
Tumorigenic: Yes
Cytogenetic Analysis: This is a hypotetraploid human cell line. The modal chromosome number was 84, occurring in 22% of cells. However, cells with chromosome counts of 86 (20%) and 87 (18%) also occurred at high frequencies. The rate of cells with higher ploidies was 6.0%.
Age: 50 years
Gender: male
Ethnicity: Caucasian
Comments: LNCaP clone FGC was isolated in 1977 by J.S. Horoszewicz, et al., from a needle aspiration biopsy of the left supraclavicular lymph node of a 50-year-old Caucasian male (blood type B+) with confirmed diagnosis of metastatic prostate carcinoma. [21889]
These cells are responsive to 5-alpha-dihydrotestosterone (growth modulation and acid phosphatase production). [23045]
The cells do not produce a uniform monolayer, but grow in clusters which should be broken apart by repeated pipetting when subcultures are prepared.
They attach only lightly to the substrate, do not become confluent and rapidly acidify the medium.
Growth is very slow.
The cells should be allowed to incubate undisturbed for the first 48 hours after subculture.
When flask cultures are shipped, the majority of the cells become detached from the flask and float in the medium.
Upon receipt, incubate the flask (in the usual position for monolayer cultures) for 24 to 48 hours to allow the cells to re-attach.
The medium can then be removed and replaced with fresh medium.
If desired, the contents of the flask can be collected, centrifuged at 300 X g for 15 minutes, resuspended in 10 ml of medium and dispensed into a single flask.
Propagation: ATCC complete growth medium: The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. 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: 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.
Maintain cultures at a cell concentration between 1 X 10(4) and 2 X 10(5) cells/cm2.
Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Medium Renewal: Twice per week
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
