LXF289 cell line细胞株细胞系 BioVector NTCC质粒载体菌种细胞基因保藏中心

LXF289 cell line细胞株细胞系 BioVector NTCC质粒载体菌种细胞基因保藏中心
Organism: Homo sapiens (human)
Ethnicity: Caucasian
Age: 62 years
Gender: Male
Tissue: Lung
Morphology: Epithelial
Cell type: Adenocarcinoma
Growth Properties: Monolayer, adherent
Description: In vitro etablished from the primary lung adenocarcinoma of a 62 year-old male.
References: Scientific proceedings. Fifth symposium of the Section of Experimental Cancer Research
(SEK) of the German Cancer Society. Heidelberg, 10-12 April, 1989. Hilbert,J.,
Goerttler,K. & Löhrke,H.. Is there an alteration of the DNA index and the cytoskeleton in
tumor cell models in comparison with xenotransplantation and in-vitro culturing? Results
of 10 human models. J Cancer Res Clin Oncol 115(Suppl. 1):Supplement: S 50, 1989.
Culture Conditions and Handling
Culture Medium: RPMI 1640 medium supplemented with 4.5g/L glucose, 2mM L-glutamine and 10% fetal
bovine serum (MG-72, CLS order number 820702).
Subculturing: Remove medium and rinse the adherent cells using PBS without calcium and
magnesium (3-5 ml PBS for T25, 5-10ml for T75 cell culture flasks). Add Accutase (1-2ml
per T25, 2.5ml per T75 cell culture flask), the cell sheet must be covered completely.
Incubate at ambiente temperature for 8-10 minutes. Carefully resuspend the cells with
medium (10 ml), centrifuge for 5 min at 300xg, resuspend cells in fresh medium and
dispense into new flasks which contain fresh medium.
Split Ratio: A ratio of 1:2 to 1:6 is recommended
Seeding density: 1x104
/ml
Fluid Renewal: 1 to 2 times weekly
Freeze Medium: CM-ACF (CLS order number 800650, 50ml)
Freezing recovery: 24-48 hrs
Sterility: Fluorescence (DAPI) test: negative; Mycoplasma specific PCR: negative; Bacteria
specific PCR: negative
Biosafety Level: 1
Special Features of the Cell Line
Tumorigenic: yes, in nude mice
Viruses: SMRV: Negative, as confirmed by Real-Time PCR
Reverse Transcriptase: negative
DNA Profile (STR): Amelogenin: X,Y
CSF1PO: 12,13
D13S317: 9,11
D16S539: 13
D5S818: 9,10
D7S820: 10,11
THO1: 6,9.3
TPOX: 11
vWA: 17,18
D3S1358: 15,18
D21S11: 30,31
D18S51: 14
Penta E: 10,20
Penta D: 10,13
D8S1179: 13
FGA: 24,25
Immunology: Cytokeratine 8, 18, positive; Desmoplakin positive; Vimentin positive

Cryopreserved cells The cells come deep-frozen shipped on dry ice. Please make sure that the vial is still
frozen.
If immediate culturing is not intended, the cryovial(s) must be stored below -150°C after
arrival.
If immediate culturing is intended, please follow these instructions:
Quickly thaw by rapid agitation in a 37°C water bath within 40-60 seconds. The water
bath should have clean water containing an antimicrobial agent. As soon as the sample
has thawed, remove the cryovial from the water bath. Note: A small ice clump should still
remain and the vial should still be cold.
From now on, all operations should be carried out under aseptic conditions.
Transfer the cryovial to a sterile flow cabinet and wipe with 70% alcohol. Carefully open
the vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of
culture medium (room temperature). Resuspend the cells carefully. Centrifuge at 300xg
for 3 min and discard the supernatant. The centrifugation step may be omitted, but in this
case the remains of the freeze medium have to be removed 24 hours later.
Resuspend the cells carefully in 10ml fresh cell culture medium and transfer them into
two T25 cell culture flasks. All further steps are described in the Subculture section.
Proliferating Cultures The cell culture flasks, 2xT25, come filled with cell culture medium.
Collect the entire medium in 2x 50 ml centrifuge tubes.
Carefully add 5 ml of cell culture medium to each of the two T25 cell culture flasks.
Control the cell morphology and confluency under the microscope.
Incubate at 37°C for a minimum of 24 hrs.
Spin down the collected medium at 300x g for 3 minutes to collect the cells which may
have detached during transit. If a cell pellet is visible, resuspend the cells in 5 ml of cell
culture medium and transfer to 1xT25 cell culture.
Incubate at 37°C for a minimum of 24 hrs.

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