L2-RYC (ATCC® CRL-2180)

Organism: Rattus norvegicus, rat  /  Tissue: embryo  /  Disease: Carcinoma

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Organism Rattus norvegicus, rat
Tissue embryo
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease Carcinoma
Age embryo; 10 days gestation
Strain Lewis
Applications
The cell line expresses parietal yolk sac endoderm characteristics in that it synthesizes basement membrane components.
Alpha fetoprotein synthesis could not be demonstrated.
Storage Conditions liquid nitrogen vapor phase
Karyotype The majority of cells had a pseudodiploid chromosome complement; approximately 10% of cells were pseudotetraploid.
Derivation
L2-RYC is a yolk sac carcinoma established in 1978 by Dr. Ulla Wewer from a yolk sac tumor induced by puncturing a Lewis rat embryo through the wall of the uterus at the 10th day of gestation.
A yolk sac carcinoma developed approximately 71/2 months later.
The L2 cell line in which only parietal carcinoma cells could be identified, was established from transplant number 14.
Genes Expressed
laminin; entactin; collagen IV; heparin sulfate proteoglycan; chondroitin sulfate proteoglycan
Cellular Products
laminin; entactin; collagen IV; heparin sulfate proteoglycan; chondroitin sulfate proteoglycan
Tumorigenic Yes
Effects
Yes, in Lewis rats
Comments
L2-RYC is a yolk sac carcinoma established in 1978 by Dr. Ulla Wewer from a yolk sac tumor induced by puncturing a Lewis rat embryo through the wall of the uterus at the 10th day of gestation.
A yolk sac carcinoma developed approximately 71/2 months later.
In the early passages of the parental cell line, designated L1, both visceral and parietal yolk sac carcinomas were present.
L1 cells were injected intraperitoneally into rats and serial transplantations were carried out.
The L2 cell line in which only parietal carcinoma cells could be identified, was established from transplant number 14.
The cell line expresses parietal yolk sac endoderm characteristics in that it synthesizes basement membrane components.
In addition, a noncartilage chondroitin sulfate proteoglycan is synthesized.
Alpha fetoprotein synthesis could not be demonstrated.
The cells must be subcultured frequently and not be allowed to become dense.
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%.
Subculturing Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.

Subcultivation Ratio: 1:5
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation

Complete growth medium described above supplemented with 5% (v/v) DMSO.  Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Name of Depositor S Thorgeirsson
Passage History
In the early passages of the parental cell line, designated L1, both visceral and parietal yolk sac carcinomas were present.
Year of Origin 1978
References

Wewer UM, et al. Heparan sulfate proteoglycans made by different basement-membrane- producing tumors have immunological and structural similarities. Differentiation 30: 61-67, 1985. PubMed: 2936642

Wewer U. Characterization of a rat yolk sac carcinoma cell line. Dev. Biol. 93: 416-421, 1982. PubMed: 7141105

Wewer U, et al. Laminin, a noncollagenous component of epithelial basement membranes synthesized by a rat yolk sac tumor. Cancer Res. 41: 1518-1524, 1981. PubMed: 7011537

. . Acta Pathol. Scand. Immunol. Sect. A, Immunol. 92: 275-283, 1984.

Oldberg A, et al. Isolation of a chondroitin sulfate proteoglycan from a rat yolk sac tumor and immunochemical demonstration of its cell surface localization. J. Biol. Chem. 256: 10847-10852, 1981. PubMed: 6793588

Lundstrom M, et al. Immunocytochemical and biochemical characterization of the Heymann nephritis antigenic complex in rat L2 yolk sac cells. Am. J. Pathol. 143: 1423-1435, 1993. PubMed: 8238258

Orlando RA, Farquhar MG. Identification of a cell line that expresses a cell surface and a soluble form of the gp330/receptor-associated protein (RAP) Heymann nephritis antigenic complex. Proc. Natl. Acad. Sci. USA 90: 4082-4086, 1993. PubMed: 8483924

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

Wewer UM, et al. Heparan sulfate proteoglycans made by different basement-membrane- producing tumors have immunological and structural similarities. Differentiation 30: 61-67, 1985. PubMed: 2936642

Wewer U. Characterization of a rat yolk sac carcinoma cell line. Dev. Biol. 93: 416-421, 1982. PubMed: 7141105

Wewer U, et al. Laminin, a noncollagenous component of epithelial basement membranes synthesized by a rat yolk sac tumor. Cancer Res. 41: 1518-1524, 1981. PubMed: 7011537

. . Acta Pathol. Scand. Immunol. Sect. A, Immunol. 92: 275-283, 1984.

Oldberg A, et al. Isolation of a chondroitin sulfate proteoglycan from a rat yolk sac tumor and immunochemical demonstration of its cell surface localization. J. Biol. Chem. 256: 10847-10852, 1981. PubMed: 6793588

Lundstrom M, et al. Immunocytochemical and biochemical characterization of the Heymann nephritis antigenic complex in rat L2 yolk sac cells. Am. J. Pathol. 143: 1423-1435, 1993. PubMed: 8238258

Orlando RA, Farquhar MG. Identification of a cell line that expresses a cell surface and a soluble form of the gp330/receptor-associated protein (RAP) Heymann nephritis antigenic complex. Proc. Natl. Acad. Sci. USA 90: 4082-4086, 1993. PubMed: 8483924

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online at http://www.cdc.gov/OD/ohs/biosfty/bmbl5/bmbl5toc.htm