769-P

€340.00*

Content: 1 cryovial
Product number: 300106
Product information "769-P"

Origin and General Characteristics


Organism                                             Human

Ethnicity                                              Caucasian

Age                                                      63

Gender                                                 Female

Tissue                                                  Kidney

Disease                                                Renal cell carcinoma

Morphology                                        Epithelial-like

Growth properties                               Adherent

Depositor                                            H. Löhrke

Cellosaurus accession ID                   CVCL_1050

Wikidata                                             Q54604830

NCBI_TaxID                                      9606

Description                                         This line was derived from a primary clear cell adenocarcinoma. The cells are globular with                                                                      indistinct borders, have a high nucleus to cytoplasm ratio and exhibit both microvilli and                                                                          desmosomes. They can be cultured in soft agar.

References                                          References are available in the product page at www.cls.shop

               

Culture Conditions and Handling

  Culture medium                     RPMI 1640, 10% FBS

Supplements

2 mM L-glutamine

Doubling time

35 hours

Subculturing

Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10 ml for T75 cell culture flasks). Add Accutase (1-2 ml per T25, 2.5 ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at ambient temperature for 8-10 minutes. Carefully resuspend the cells with medium (10 ml), centrifuge for 3 min at 300 g, resuspend cells in fresh medium and dispense into new flasks which contain fresh medium.

Split ratio

A ratio of 1:4 to 1:12 is recommended

Seeding density

3 x 10^4 cells/cm^2 will result in a confluent monolayer within 4 days.

Fluid renewal

Every 2 to 3 days

Freeze medium

CM-1 (CLS catalog number 800150)

Freeze medium

After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 48 hours.

Sterility

Plasmotest: negative; Mycoplasma specific PCR: negative        

 

Special Features of the Cell Line


Tumorigenic

Yes, in immuno-suppressed hamsters and nude mice

Viruses

SMRV: Negative, as confirmed by Real-Time PCR

Karyotype

Hypodiploid; modal number = 45; a large submetacentric chromosome

was present in all cells.

 

 

DNA profile (STR)

 

Amelogenin

CSF1PO

X,X

11,12

D18S51

Penta E

14,17

7,18

 

D13S317

10,14

Penta D

12,16

 

D16S539

9,13

D8S1179

12,16

 

D5S818

12

FGA

20,22

 

D7S820

10,11

D1S1656

n.a.

 

TH01

6,9.3

D6S1043

n.a.

 

TPOX

8,11

D2S1338

n.a.

 

vWA

18

D12S391

n.a.

 

D3S1358

16

D19S433

n.a.

 

D21S11

28,30

 

 

 

 

 

 

 

HLA-typing

A*

n.a., n.a.

DQB1*

n.a., n.a.

 

B*

n.a., n.a.

DPA1*

n.a., n.a.

 

C*

n.a., n.a.

DPB1*

n.a., n.a.

 

DRB1*

n.a., n.a.

E

n.a., n.a.

 

DRB3*

n.a., n.a.

G

n.a., n.a.

 

DRB4*

n.a., n.a.

F

n.a., n.a.

 

DRB5*

n.a., n.a.

MR-1

n.a., n.a.

 

DQA1*

n.a., n.a.

 

 

 

 

 

 

 

 

 

 

 

 

Recommendations for handling of cells following delivery

image

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 must be stored below -150°C after arrival. If immediate culturing is intended, please follow the below 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. 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 300 x g 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 10 ml 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 2 x 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 hours. Spin down the collected medium at 300 x 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 1 x T25 cell culture.

Incubate at 37 °C for a minimum of 24 hours.

 

 

Legal & Documentation

image

Safety precautions

If the cryovial is planned to be stored in liquid nitrogen and to be thawed in the future, special safety precautions should be followed. Protective gloves and clothing should be used and a facemask or safety goggles must be worn when transferring frozen samples into or removing from the liquid nitrogen tank. The removal of a cryovial from liquid nitrogen may result in the explosion of the frozen vial creating flying fragments.

Warranty

CLS warrants for a high cell viability and culture performance only if the products are stored and cultured according to the information described in this product sheet.

Certificate of Analysis

The certificate of analysis can be requested by email at info@cls.shop for each batch. Please indicate the lot number of your product in the email.

Disclaimer

This product is intended for laboratory research use only. It is strictly prohibited in any case to use the cells, their progeny, unmodified derivatives and modifications in human or animal subjects for therapeutic, diagnostic or prophylactic purposes. All regulations as described in the Material Transfer Agreement or Supply Agreement concluded with the order apply.


 

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Required products

Freeze Medium CM-1
- Ready-to-use and easy-to-use- Suitable for long-term cryopreservation- Tested with more than 400 cell lines- Stable for more than 6 monthsCM-1 represents a cryopreservation medium developed by CLS enabling the long-time cryopreservation of Human and Animal cells and cell lines at -196°C. For more than 15 years it has been used successfully at CLS, therefore consistant results with respect to cell viability following the thawing procedure are at hand. 

Variants from €29.00*
€39.00*
RPMI 1640
RPMI-1640 represents an excellent cell culture medium for human and animal lymphocytes, either normal or neoplastic. 

€18.00*
Accutase
Accutase Cell Detachment solution is a ready-to-use cell detachment solution of proteolytic and collagenolytic enzymes. Useful for the routine detachment of cells from standard tissue culture plastic ware and adhesion coated plastic ware, and polymers. Accutase performs exceptionally well in detaching cells for the analysis of cell surface markers, virus growth assays, quiescence assays by serum starvation, transformation assays by oncogene transfection, neural crest cell migration assays, cell proliferation, cell haptotaxis, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.Accutase does not contain mammalian or bacterial derived products.  

€43.95*
FBS
- Fetal Bovine Serum- Cell culture grade- EU-compliant- Origin: South AmericaWe recommend sterile filtration through 0.22 µm filter. 

Variants from €25.20*
€219.00*
PBS
Phosphate buffered solutionSterile, applicable in cell culture 

€7.00*

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